1st Chilean German Workshop

Developmental Biology and Regenerative Medicine

From tissue generation to re-generation
1 4 March 2013, Santiago de Chile
P. Universidad Catlica de Chile, Casa Central, Alameda 324, piso 2
Topics
Organ and Tissue Development Wound Healing and Regeneration
Tissue Engineering Stem Cells - Inflammation Biomaterials -
Model Organisms Surgical Research
Speakers
Miguel Allende / J.Ignacio Arias / Marcelo Campos / Julian Dye / Miguel Allende / J.Ignacio Arias / Marcelo Campos / Julian Dye /
Toms Egaa / Alberto Fierro / Fernando Fierro / Yves Harder /
Claudio Hetz / Gudrun Kausel / Maroun Khoury / Juan Larran /
Sergio Lavandero / Hans-Gnther Machens / Martn Montecino /
Andreas Nssler / Vernica Palma / Norbert Pallua /
Sebastin San Martn / Peter Vogt
Organized by:
FONDAP Center for Genome Regulation, CGR
Miguel Allende, Universidad de Chile
Toms Egaa, Technische Universitt Mnchen
INFORMATION: www.genomacrg.cl workshop2013@genomacrg.cl
!"##$%&'( )*+,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,!#$-.$%'( )*+
1

WLLCCML WCkDS
llrsL Cerman-Chllean Workshop on uevelopmenLal 8lology and
8egeneraLlve Medlclne. lrom Llssue generaLlon Lo re-generaLlon.


SanLlago de Chlle
March 1Lh Lo 4Lh, 2013

uear Colleagues,

ueveloplng lnnovaLlve and effecLlve sLraLegles ln 8egeneraLlve Medlclne requlres
cuLLlng-edge research Lechnologles and Lhe esLabllshmenL of new sLraLeglc
lnLerdlsclpllnary parLnershlps worldwlde.
Almlng Lo provlde a plaLform for esLabllshlng long-Lerm sclenLlflc and academlc
collaboraLlons beLween Chllean and Cerman research lnsLlLuLlons, we would llke Lo
welcome you Lo Lhe I|rst Ch||ean-German Workshop on Deve|opmenta| 8|o|ogy and
kegenerat|ve Med|c|ne.
We hope Lhls meeLlng represenLs a unlque opporLunlLy for sharlng ldeas and you en[oy
your Llme ln Chlle.

Slncerely,



M|gue| A||ende 1oms Lgaa
unlversldad de Chlle 1echnlsche unlverslLaL Munchen

2


Sponsored by:
Supported by:
3

lCnuA CenLer for Cenome 8egulaLlon



Cur cenLer conslsLs of an lnLerdlsclpllnary nucleus of sclenLlsLs, lncludlng blologlsLs and
maLhemaLlclans of Lhree lmporLanL naLlonal unlverslLles, uolvetslJoJ Je cblle, lootlflclo
uolvetslJoJ cotllco and uolvetslJoJ AoJts 8ello.
1hls Leam ls wldely experlenced ln Lhe use of model organlsms, ln addlLlon Lo belng
recognlzed experLs ln Lhe use of advanced genomlc Lechnologles, blolnformaLlcs and
SysLems 8lology.
We look Lo Lake advanLage of Lhe excepLlonal quallLy of fundamenLal sclence carrled ouL
ln our academlc labs, wlLh Lhe purpose of provldlng resources LhaL wlll lead Lo Lhe
developmenL of lnnovaLlve approaches Lo bloLechnologlcal problems LhaL Lhe counLry
has deLermlned are sLraLeglc for lLs developmenL.
1he cenLer provldes, Lo Lhe Chllean and LaLln Amerlcan sclenLlflc communlLy, Lhe
lnfrasLrucLure and research aLmosphere LhaL wlll sLlmulaLe fuLure developmenLs ln flelds
LhaL range from cllnlcal dlagnosls Lo lmprovemenL of Lhe crops and susLalnable use of
naLural resources.
Cur alm for Lhe comlng years ls Lo esLabllsh Lhe CC8 as a naLlonal reference cenLer ln
Lhe area of genomlcs LhaL approaches a varleLy of blologlcal quesLlons and responds Lo
Lhe producLlve challenges faced by Lhe counLry.


4

kCGkAM
IkIDA 01.03.2013

13:00-16:43

8eglsLraLlon
16:43 Welcome words
M|gue| A||ende, ulrecLor CenLer for Cenome 8egulaLlon
17:00 Welcome words
Ambassador Gabr|e| kodr|guez, Chllean MlnlsLry of lorelgn Affalrs
17:30 keynoLe lecLure:
Iu||an Dye, SclenLlflc ulrecLor 8Al1 lnsLlLuLe, uk
1be 5mott Mottlx
1M
syotbetlc Jetmol scoffolJ fot sklo tecoosttoctloo
18:1S Cockta||


3


SA1UkDA 02.03.2013

Cha|rs: Iu||an Dye & Vern|ca a|ma
09:00 ves narder, 1echnlsche unlverslLaL Munchen, CL8MAn?
1lssoe ptecooJltlooloq lo plostlc sotqety. ltom beocb to beJslJe
09:13 Iernando I|erro, unlverslLy of Callfornla aL uavls, uSA
use of meseocbymol stem cells os bollJloq blocks ooJ otcbltects
09:40 Marce|o Campos, unlversldad de Chlle, CPlLL
colloqeos lo bomoo tototot coff tlssoes oftet sbock wove tteotmeot.
A komoo ooJ sotfoce-eobooceJ komoo scottetloq stoJy
09:33 eter Vogt, Pannover Medlcal School, CL8MAn?
18A
10:30 Coffee break

Cha|rs: Sebast|n San Mart|n & Iuan Larra|n
11:00 Vern|ca a|ma, unlversldad de Chlle, CPlLL
Neoqeolo- 1 moJolotes ooqloqeoesls loJoceJ by oettlo-1 ptoJoceJ by bomoo
wbottoos jelly meseocbymol stem cells (wI-M5c)
11:13 M|gue| A||ende, unlversldad de Chlle, CPlLL
1be 2ebtoflsb os o televoot moJel fot vettebtoteoeotol teqeoetotloo
11:40 A|berto I|erro, Cllnlca Las Condes, CPlLL
loltlol steps towotJs oo oJoptlve tbetopy wltb teqolototy 1 cells
11:33 osLer prevlews (2 mlnuLes each)
12:30-14:00 Lunch at "GAM"

14:00 oster sess|on and coffee
Cha|rs: Iernando I|erro & ves narder
16:00 Norbert a||ua, 8W1P Aachen unlverslLy PosplLal, CL8MAn?
5tem cells ooJ oJlpoqeoesls - lo vltto ooJ lo vlvo 1lssoe oqloeetloq
16:23 Serg|o Lavandero, unlversldad de Chlle, CPlLL
now to tepolt o btokeo beott? !
16:30 Andreas Nss|er, Lberhard karls unlverslLaL 1ublngen, CL8MAn?.
1be tole of tbe llvet lo booe Jomoqe. two moJels to stoJy bepotlc OsteoJysttopby
17:13 Gudrun kause|, unlversldad AusLral, CPlLL
Cetmoo keseotcb loooJotloo (ulC) ooJ ptoqtoms to soppott lotetootloool
coopetotloo.
20:00 Speakers d|nner

6

SUNDA 03.03.2013

Cha|rs: Marce|o Campos & Norbert a||ua
09:00 Maroun khoury, unlversldad de Los Andes, CPlLL
nomo moscolos, mote tboo jost o moose wltb bomoo blooJ?
09:13 I. Ignac|o Ar|as, unlversldad de Chlle, CPlLL
1typooosomoctozl coltetlcollo, o potoslte ptotelo tbot beols
09:40 nans-Gnther Machens, 1echnlsche unlverslLaL Munchen, CL8MAn?
cllolcol cbolleoqes ooJ teseotcb oppottooltles lo llostlc 5otqety
10:1S- 11:00 Coffee break

Cha|rs: eter Vogt & Serg|o Lavandero
11:00 Iuan Larra|n, onLlflcla unlversldad CaLllca, CPlLL
5plool cotJ keqeoetotloo lo \eoopos!
11:23 Andreas Nss|er, Lberhard karls unlverslLaL 1ublngen, CL8MAn?
1be tole of oqloq lo oJlpose-JetlveJ stem cells ooJ tbe coosepoeoces fot
tbelt lo-vltto ooJ lo-vlvo ose
11:30 1oms Lgaa, 1echnlsche unlverslLaL Munchen, CL8MAn?
uevelopmeot of blooctlve scoffolJs fot tlssoe eoqloeetloq ooJ teqeoetotloo
12:30-14:00 Lunch at yourse|f

Cha|rs: nans-Gnther Machens & A|berto I|erro
14:00 Gudrun kause|, unlversldad AusLral, CPlLL
Metol sttess oo pltoltoty foctots lo cyptlooscotplo
14:13 Iu||an Dye, 8Al1 lnsLlLuLe, uk
1be potbwoy fot cllolcol ttooslotloo of 5mott Mottlx
1M
os oo exomple 1lssoe
5coffolJ ptoJoct
14:40 Sebast|n San Mart|n, unlversldad de valparalso, CPlLL
lloceotol stem cell, ftom boslc teseotcb to cllolcol oppllcotloos!
13:10 - 13:23 N|ko|aos apadopu|os, 1echnlsche unlverslLaL Munchen, CL8MAn?
1owotJ o mooofoctotloq ptocess of bomoo oJlpose sttomol cells fot cell
tbetopy. ltellmlooty tesolts
16:30 Speaker's soc|a| event:
1r|p to "INDCMI1A" v|neyard, Casab|anca Va||ey

7

MCNDA 04.03.2013

Cha|rs: Andreas Nss|er & Maroun khoury
09:30 Mart|n Montec|no, unlversldad Andres 8ello, CPlLL
plqeoetlc coottol of exptessloo of booe-mostet teqolotots Jotloq osteoblost
lloeoqe commltmeot!
09:33 ves narder, 1echnlsche unlverslLaL Munchen, CL8MAn?
Aotoloqoos fot qtoftloq to cottect osymmetty followloq tecoosttoctlve
sotqety of tbe bteost. 1be lofloeoce of smokloq ooJ toJlotbetopy.
10:10 C|aud|o netz, unlversldad de Chlle, CPlLL
ltotelo folJloq sttess lo oeotoJeqeoetotlve Jlseose. oo lotetploy
betweeo k sttess ooJ ootopboqy
10:4S Coffee break

11:30 - 13:00 kound tab|e d|scuss|on
Gonza|o Arenas, CCnlC?1 (Chllean 8esearch Councll)
Oppottooltles fot tbe lotetootlooollzotloo of cblleoo scleoce
13:00 I|na| remarks



8

kLNC1L LLC1UkL

1he Smart Matr|x Synthet|c Derma| Scaffo|d for sk|n reconstruct|on
!ullan l uye
ulrecLor of 8esearch, 8Al1 lnsLlLuLe, MounL vernon PosplLal, London, uk
8ecaplLuallng elemenLs of developmenL ln human wound reconsLrucLlon ls an lmporLanL
yeL eluslve goal. ln splLe of Lhree decades of arLlflclal skln sLraLegles Lhere ls sLlll an
urgenL un-meL cllnlcal need for effecLlve expedlenL sLraLegles for reconsLrucLlon of full-
Lhlckness skln-loss,. SmarL MaLrlx` (SM) ls a cross-llnked flbrln-alglnaLe porous
composlLe developed as a pro-vascular synLheLlc dermal scaffold. 1he lmpeLus for lLs
developmenL arose from llmlLed cllnlcal success of hosL lnLegraLlon & long-Lerm
ouLcomes of some prevlous blomaLerlals. rlor experlence has shown LhaL Lhe neo-
vascularlsaLlon raLe could llmlL boLh speed and ulLlmaLe success or fallure (from lack of
lnLegraLlon, fluld accumulaLlon, lnflammaLlon, lnfecLlon) of arLlflclal skln maLerlals..
1he deslgn of SM focussed on creaLlng a new blomaLerlal wlLh Lhe pro-anglogenlc
properLles of flbrln. LngrafLlng proLoLype SM ln a full-Lhlckness porclne skln-exclslon
wound rlng model demonsLraLed deep cellular lngress wlLhln 7 days wlLh a
neovascularlsaLlon process resembllng vasculogenesls, ln addlLlon Lo wound bed
anglogenesls. 8y week 3, Lhe scaffold/Llssue conLracLlon was <20 and resulLlng neo-
dermal Llssue showed dlmlnlshlng oSM-acLln poslLlve flbroblasLs and oSM-acLln poslLlve
caplllarles.
1he blomaLerlal sLrucLure was developed prlmarlly Lo maxlmlse lnLegraLlon raLe.
CpLlmlsed SM demonsLraLes compleLe cellularlsaLlon and neo-vascularlsaLlon
LhroughouL Lhe scaffold by day 7. lmporLanLly, Lhe cellular and vascular lnLegraLlon of
SmarL MaLrlx ls rapld enough for lnLegraLlon of 0.13 mm spllL-Lhlckness skln grafL (slngle
procedure full-Lhlckness reconsLrucLlon). 8esulLs aL week 6 demonsLraLe Lhe neodermls
resembles surroundlng skln boLh by palpaLlon and cellular organlsaLlon, whlch lacks
obvlous adnexal sLrucLures buL has feaLures conslsLenL wlLh regeneraLed Llssue.
1hese precllnlcal resulLs suggesL cllnlcal beneflLs from acceleraLed lnLegraLlon,
mlnlmlsed compllcaLlons, and benlgn ouLcome.

9









A8S1kAC1S CkAL kLSLN1A1ICNS
(A|phabet|c order)

10

1he zebraf|sh as a re|evant mode| for vertebrate neura| regenerat|on
Mlguel L Allende, 8osarlo vlllegas, Marla Laura Cecl, !ose Moya, Slmn Carrlllo, nlcole
8eynaerL, Cscar ena, Marlo Snchez, Mar[orle Alvarez, Camlla Mardones, CrlsLlan
undurraga.

lCnuA CenLer for Cenome 8egulaLlon. laculLad de Clenclas, unlversldad de Chlle.

lor Lhe pasL LwenLy years, Lhe zebraflsh model has been growlng ln lmporLance for baslc
sLudles ln developmenLal geneLlcs. More recenLly, lL has also become a conLrlbuLlng
force ln dlsease models and drug screens, Lransformlng lL lnLo a new player ln precllnlcal
sLudles. A sequenced genome and advanced gene manlpulaLlon Lechnologles are
complemenLed by lLs aLLracLlve embryos and larvae: Lhelr Lransparency permlLs ln vlvo
monlLorlng of cells and gene expresslon ln real Llme. lurLhermore, ln Lhese anlmals, loss
of many cell Lypes and organs ls followed by robusL regeneraLlon, a dlsLlngulshlng
feaLure of LeleosLs. 1hus, Lhe zebraflsh ls an ldeal sysLem for regeneraLlon research and
for deLecLlng molecules LhaL lnfluence Lhe regeneraLlve capaclLy of dlfferenL organs. We
are analyzlng Lhe cellular and molecular mechanlsms LhaL lmpacL on Lhe ablllLy of
perlpheral neurons and sensory cells Lo regeneraLe afLer damage or loss. We can
damage axons by preclse laser or mechanlcal secLlonlng, whlle exLernal
mechanosensory cells can be ablaLed chemlcally. ln boLh cases, flsh show a remarkable
ablllLy Lo rebulld sLrucLures ouL of exlsLlng componenLs. We have shown LhaL sensory
neuron axons efflclenLly regrow from remalnlng somas afLer axoLomy. 1he growLh of
axons afLer ln[ury depends on dlfferenL cell Lypes lncludlng Schwann cells and lnnaLe
lmmune cells. Mechanosensory cells are newly generaLed from a populaLlon of
progenlLor cells, resldlng aL Lhe base of Lhe organs Lhey lnhablL. ln Lhls process, we have
also observed parLlclpaLlon of cells of Lhe lmmune sysLem, Lhough Lhelr exacL role ln
promoLlng or lnhlblLlng regeneraLlon remalns Lo be deLermlned. As many of our sLudles
are carrled ouL uslng larval sLage anlmals, Lhe analysls can be done ln hlgh-LhroughpuL
formaL. 1hus, molecule screens LhaL alm Lo uncover blologlcal acLlvlLles relaLed Lo
regeneraLlon and lmmunlLy are posslble. llnally, we have begun sLudles ln whlch we wlll
dlssecL Lhe lmpacL of oxldaLlve sLress and age on Lhe speed and efflclency of
regeneraLlon ln Lhese dlfferenL organ sysLems. Cur goal, ls Lo deLecL paLhways LhaL are
affecLed under Lhese condlLlons whlch may offer LargeLs for cllnlcal lnLervenLlon.

lundlng: lCnuA 13090007, lCnuLC?1 1110273.


11

1rypanosama cruz| Ca|ret|cu||n, a paras|te prote|n that hea|s

!. lgnaclo Arlas
1
, Caroll Sepulveda
1
, ChrlsLopher PamllLon-WesL
1
, lsmael Maldonado
2
and
ArLuro lerrelra
2

1
uepLo. Clenclas Cllnlcas, laculLad de Clenclas veLerlnarlas, unlversldad de Chlle,
SanLlago, Chlle.
2
lC8M, laculLad de Medlclna, unlversldad de Chlle, SanLlago, Chlle.

CalreLlculln (C81) ls a ublqulLous proLeln wlLh a remarkable conservaLlon ln all specles, ls
found malnly ln Lhe endoplasmlc reLlculum (L8), where lL ls fundamenLal for Lhe proper
foldlng and LransporLaLlon of proLelns across Lhe L8. lL has been descrlbed varlous
exLracellular funcLlons for Lhls proLeln, lncludlng lnducLlon of phagocyLosls of apopLoLlc
cells, as well as promoLlon of concenLraLlon-dependenL mlgraLlon keraLlnocyLes,
flbroblasLs, macrophages and monocyLes, sLlmulaLlon of Lhe re-eplLhellallzaLlon process
Lhrough keraLlnocyLe and flbroblasLs cell prollferaLlon, whlch ls an essenLlal processes
for cuLaneous wound heallng. Accordlng Lo Lhese funcLlons, ls dlfferenL effecLs were
evaluaLed ln Lhe wound heallng process ln skln, by subcuLaneously admlnlsLraLed
recomblnanL human (rPuC81) and 1rypanosoma cruzl (r1cC81) C81 ln varlous
concenLraLlons way Lo compare Lhe effecLlveness of boLh proLelns ln Lhe percenLage of
re-eplLhellallzaLlon, cell prollferaLlon (measured as numbers of cells), deep dermal and
epldermal depLh, boLh aL 3 and 10 days posL-lnLervenLlon. 1he besL resulLs ln heallng of
skln wounds ln raLs were obLalned by admlnlsLerlng r1cC81 ln lower concenLraLlons
(2,300 Llmes less concenLraLed Lhan Lhe hlgh dose) and rPuC81 of hlgher concenLraLlon.


12

Co||agens |n human rotator cuff t|ssues after shock wave treatment. A kaman and
surface-enhanced kaman scatter|ng study

!.!. Crcamo
1
, 8. L. Clavl[o
2
, M.8. 8ranes
2
and M.M. Campos-valleLLe
2
.

1
unlversldad de 1arapac. upLo. de AnLropologla, Arlca, Chlle.
2
LaboraLory of vlbraLlonal
SpecLroscopy, laculLad de Clenclas, unlversldad de Chlle, Casllla 633, SanLlago, Chlle.

lmporLanL lmprovemenLs of chronlc LendlnopaLhles Lendons dlseases are seen afLer
shockwave (SW) LreaLmenL. neo-anglogenesls sLlmulaLlon and hypercellularlzaLlon are
Lhe resulL of shorL Llme perlods of LreaLmenL. 1he presenL work provldes a flrsL
approach from a molecular vlew polnL of Lhese blo-processes, mosL llkely assoclaLed Lo
sLrucLural aspecLs resulLlng from blochemlcal changes orlglnaLed by Lhe SW.
lmmunohlsLochemlcal daLa lndlcaLe LhaL collagen areas ln Lhe Llssues are lnfluenced Lhe
mosL by SW. AddlLlonal presence of collagen Lypes l and lll by SW LreaLmenL ls lnferred
from an observed lncreaslng of Lhe Llssues LlncLorlal properLles. 1he Lools selecLed for
Lhe sLudy are 8aman specLroscopy and Lhe ulLra senslLlve Lechnlque surface-enhanced
8aman scaLLerlng (SL8S). Pere we exLracL lnformaLlon from specLra of 32 blopsles of
human Lendon Llssues on Ag nanoparLlcles before and afLer SW LreaLmenL. 1he specLral
lnformaLlon ls analyzed on Lhe basls of 8aman and SL8S daLa of collagens Lype l and lll
and lLs mosL abundanL amlno acld componenLs. SL8S specLra on Lhe collagen area of
Llssues reveal Lhe presence of characLerlsLlc modes relaLed malnly Lo amlno aclds. lL has
been found LhaL Lhe maln dlfferences beLween Llssue samples are correlaLed wlLh
collagen sLrucLural conformaLlonal aspecLs.


13

1he pathway for c||n|ca| trans|at|on of Smart Matr|x(1M) as an examp|e 1|ssue Scaffo|d
product.
!ullan l uye
8Al1 lnsLlLuLe, MounL vernon PosplLal, London, uk.
1he LranslaLlonal developmenL of SmarL MaLrlx` (SM) lnvolves a breadLh of experLlse Lo
address sclenLlflc challenges. SM, a flbrln-alglnaLe based porous blomaLerlal, was
developed as a pro-vascular synLheLlc dermal scaffold, lnLended for reconsLrucLlon of
full-Lhlckness skln loss wounds. 1ranslaLlon of Lhe ldea lnLo a LherapeuLlc producL
uncovers a broad conLexL ln whlch Lhe sclenLlflc lssues need Lo be consldered. 1hls
declslons made Lo deflne Lhe paLhway of LranslaLlon crlLlcally lnfluence Lhe pro[ecL's
cllnlcal developmenL sLraLegy.
An lmporLanL lssue ls how Lo deflne Lhe un-meL cllnlcal need, ln Lhe conLexL of currenL
sLandard of pracLlce and undersLandlng of Lhe paLhophyslology. 1he lnlLlal cllnlcal
problem ls Lhe lnLegraLlon process, especlally focusslng on vascularlsaLlon, whlch ls
crlLlcal. Powever, Lhe long-Lerm ouLcome ls of ulLlmaLely cllnlcal lmporLance. 8oLh
requlre sulLable pre-cllnlcal models ln whlch Lo derlve evldence. AppreclaLlon of Lhe
array of acuLe and paLhologlcal causes of, and responses Lo, skln loss wlll gulde Lhe
developmenL of evldence for cllnlcal appllcaLlon. 1hls wlll seL Lhe parLlcular LherapeuLlc
sLraLegy Lo focus Lhe furLher developmenL of Lhe producL. ln Lhe case of synLheLlc skln
Lhls ls lmporLanL as Lhere are several dlsLlncL sLraLegles whlch have been adopLed ln
prevlous and currenL producLs. SlgnlflcanL dlfflculLles around creaLlon of relevanL ln-vlvo
models of human chronlc dlsease sLaLes seL llmlLs Lo developlng evldence for Lhe range
of appllcablllLy and clalms for Lhe producL.
1he regulaLory classlflcaLlon and l sLraLegy also deLermlne Lhe LranslaLlonal paLhway.
1he manufacLurlng and commerclallsaLlon framework requlres Lhe developers Lo
undersLand Lhe regulaLory process and provlde a whlch demonsLrably compllanL
operaLlonal sLrucLure. 1hese key conslderaLlons and declslons fundamenLally lnfluence
Lhe producL's cllnlcal developmenL.

14

Deve|opment of 8|oact|ve Scaffo|ds for 1|ssue Lng|neer|ng and kegenerat|on
!. 1oms Lgana
uepL. lasLlc and Pand Surgery, kllnlkum rechLs der lsar, 1u Munchen & londap CenLer
for Cenome 8egulaLlon, laculLad de Clenclas, unlversldad de Chlle.
uurlng wound heallng lsolaLed cells cannoL resLore Lhelr respecLlve Llssue archlLecLure
because Lhey lack a LemplaLe Lo gulde Lhelr growLh. Such LemplaLes are known as
scaffolds and Lhelr developmenL ls a ma[or Loplc ln Lhe fleld of regeneraLlve medlclne.
AlLhough huge efforLs have been made Lo develop new Lechnologles and blomaLerlals,
auLologous Llssue LransplanLaLlon ls sLlll Lhe gold sLandard procedure for LreaLlng Llssue
defecLs. 1he maln drawback of Lhe scaffolds ls Lhe cllnlcal fallure due Lo delays ln
vascularlsaLlon. As a consequence of such low vascularlsaLlon, scaffolds are noL properly
perfused wlLh cells, nuLrlenLs and oxygen. ln order Lo lmproved vascularlsaLlon we have
comblned Lhe use of collagen scaffolds for dermal regeneraLlon wlLh sLem cells obLalned
from dlfferenL Llssue sources. 8esulLs lo vlvo shows LhaL vascularlsaLlon of seeded
scaffolds was slgnlflcanLly lmproved by Lhe presence of Lhe cells. lo vltto resulLs suggesL
LhaL such lnducLlon ln vascularlsaLlon ls due paracrlne effecLs raLher Lhan sLem cell
dlfferenLlaLlon.

13

In|t|a| Steps 1owards an Adopt|ve 1herapy w|th kegu|atory 1 Ce||s

Candla L
4
, Covlan C
2
, 8odrlguez lS
2
, Morales !
4
, 8osemblaLL M
2,3,3,
llerro !A
1,4


1
laculLad de Medlclna, unlversldad de Chlle.
2
unlversldad Andres 8ello.
3
lundacln
Clencla para la vlda.
4
unldad de 1rasplanLe, Cllnlca Las Condes.
3
laculLad de Clenclas,
unlversldad de Chlle.

AdmlnlsLraLlon of regulaLory 1 cells (1reg) emerges as a sulLable sLraLegy Lo modulaLe
auLolmmunlLy as well as bone marrow and organ LransplanLaLlon. lo vlvo and ex
vlvosLraLegles are envlsaged. revlous work performed ln mlce has shown Lhe feaslblllLy
of generaLlng and expandlng ex vlvo 1reg from nalve 1 cells (1n) ln Lhe seLLlng of
experlmenLal LransplanLaLlon. ln Lhe presenL sLudy we alm Lo generaLe and expand
polyclonal human 1reg ex vlvo as a flrsL sLep Lo modulaLe Lhe lmmune response.
erlpheral blood mononuclear cells obLalned from healLhy volunLeers were enrlched
Lhrough denslLy gradlenLs, 1n lsolaLlon was performed by negaLlve selecLlon uLlllzlng a
cockLall of bloLln con[ugaLed anLlbodles and anLl-bloLln Mlcro8eads. 1n were
polyclonally sLlmulaLed wlLh anLl-Cu3 and anLl-Cu-28 con[ugaLed beads, and LhereafLer
culLlvaLed ln medla supplemenLed wlLh lL-2, 1Cl-beLa and all-Lrans reLlnolc acld (A18A)
aL dlfferenL concenLraLlons. 8esulLs show LhaL 1Cl-beLa ls necessary for Lhe generaLlon
of 1reg, however a conLamlnaLlng populaLlon of convenLlonal 1 cells ls also
observed. A18A ln con[uncLlon wlLh lL-2 and 1Cl-beLa lncrease furLher Lhe number of
1reg. AfLer characLerlzaLlon, furLher lsolaLlon of 1reg wlLh cell sorLer allows purlLy above
99 .

SupporLed by londecyL 1120731 and Cllnlca Las Condes.



16

Use of Mesenchyma| Stem Ce||s as bu||d|ng b|ocks and arch|tects
lernando llerro
unlverslLy of Callfornla aL uavls, uSA.
Mesenchymal sLem cells/mulLlpoLenL bone marrow sLromal cells (MSC) have become
prlme candldaLes for dlfferenL cellular Lheraples, because of Lhelr slmple lsolaLlon, greaL
expanslon poLenLlal and low lmmunogenlclLy. Whlle many appllcaLlons rely on Lhe
secreLlon of Lrophlc facLors by MSC (MSC as archlLecLs, modulaLlng anglogenesls,
lmmune response, eLc.),when used for bone, carLllage and Lendon repalr, MSC
dlfferenLlaLe lnLo Lhe respecLlve cell LypesLo dlrecLly replace Lhe damaged Llssue (acLlng
as bulldlng blocks). 1hls presenLaLlon wlll focus on our maln pro[ecLs lnvolvlng MSC ln
boLh baslc and applled sclences. llrsL, we have focused on Lhe role of mlcro8nAs ln
MSC, where we show Lhelr essenLlal funcLlon conLrolllng cell prollferaLlon and
dlfferenLlaLlon. Second, we sLudled Lhe effecL of pre-condlLlonlng MSC under low
oxygen levels, whlch shows poLenL anLl-apopLoLlc effecLs. llnally, we are conducLlng Lwo
hase l cllnlcal Lrlals, one Lo LreaL PunLlngLon's ulsease (Pu) and one Lo LreaL CrlLlcal
Llmb lschemla (CLl).



17

1|ssue precond|t|on|ng |n p|ast|c surgery: from bench to beds|de
?ves Parder, larld 8ezaelan, uanlel Schmauss, 1oms Lgana, Pans-CunLher Machens


ueparLmenL for lasLlc Surgery and Pand Surgery, kllnlkum rechLs der lsar, 1echnlsche
unlverslLaL Munchen, Cermany.

Wlde dlssecLlons and Lransfer of large Llssue compounds (flaps) - nowadays used on a
regular base Lo reshape bodles respecLlvely reconsLrucL defecLs - are dependenL on
sufflclenL perfuslon parLlcularly wlLhln Lhe randomlzed areas dlsLanL Lo Lhe flap's
pedlcle(s) (ln-flow). Accordlngly, lnadequaLe perfuslon evenLually deLermlnes morbldlLy,
whlch ln Lurn can be reduced followlng ,surglcal delay, l.e. Lhe sLepwlse dlssecLlon of a
flap ln order Lo lnduce hypoxla and so arLerlogenesls and anglogenesls. 1hls lnvaslve and
long lasLlng repeLlLlve procedure can be replaced by Llssue precondlLlonlng LhaL ls able
Lo lncrease lschemlc Lolerance of Llssues and/or malnLaln mlcrovascular perfuslon wlLhln
Lhe Llssues aL rlsk. recondlLlonlng ls assoclaLed wlLh an lncrease of proLelns such as
heaL shock proLelns LhaL can be lnduced followlng Lhe appllcaLlon of unspeclflc
supraphyslologlcal sLressors (e.g. LranslenL lschemla, local heaL, mechanlcal sLress,
plelLroplc molecules). uslng an lo vlvo model of crlLlcal lschemla of Lhe skln, we
demonsLraLe our experlence wlLh local and sysLemlc precondlLlonlng and hlghllghL our
flrsL cllnlcal experlences. 1hereby, we focus on slmple and safe appllcablllLy, efflcacy
and cosL-effecLlveness.

18

Auto|ogous fat graft|ng to correct asymmetry fo||ow|ng reconstruct|ve surgery of the
breast: the |nf|uence of smok|ng and rad|otherapy.
?ves Parder, uanlel Muller, Maxlmlllan Lder, Laszlo kovacs, Allan A. Allan, !ascha Lll,
Pans-CunLher Machens, !an-1horsLen SchanLz

ueparLmenL for lasLlc Surgery and Pand Surgery, kllnlkum rechLs der lsar, 1echnlsche
unlverslLaL Munchen, Cermany.

AuLologous faL grafLlng from processed llpoasplraLe (llpofllllng") ls lncreaslngly used Lo
correcL asymmeLry of volume and shape. 8ecenL sLudles ldlcaLe LhaL llpofllllng of Lhe
breasL ls assoclaLed wlLh a low morbldlLy, an almosL unaffecLed radlologlcal follow-up
and a predlcLable resorpLlon raLe. nelLher Lhe recurrence raLe nor Lhe Je oovo
formaLlon of breasL cancer have been proven so far. 1he sLudy alm was Lo evaluaLe
safeLy and predlcaLblllLy of llpofllllng Lo Lhe reconsLrucLed breasL Laklng lnLo accounL
acLlve smoklng and radlaLlon of Lhe Lhoraclc wall/breasL. We reporL a consecuLlve
analysls of 77 llpofllllng-procedures ln 39 paLlenLs over a 27-monLh perlod lncludlng
paLlenL demographlcs and rlsk facLors. lollow-up conslsLed of a quesLlonnalre,
phoLography and volumeLry (3-u surface scan) before and afLer surgery and was
correlaLed Lo lo vltto analysls of Lhe llpoasplraLe.
Mean ln[ecLlon volume: 149ml (30ml-490ml). Mean resorpLlon raLe aL 18 monLhs: 41
(37-31) wlLh a Lendency Lo lncreased faL resorpLlon ln smokers and afLer
radloLherapy. erloperaLlve morbldlLy: none. osLoperaLlve morbldlLy: 3. aLlenL
saLlsfacLlon aL 12 monLhs (excellenL or good wlLh regard Lo conslsLency, shape and
symmeLry): 96. AuLologous faL grafLlng for reflnemenL-surgery afLer reconsLrucLlve
breasL surgery ls safe. AcLlve smoklng and lrradlaLed skln are assoclaLed wlLh an
lncreased resorpLlon raLe of Lhe faL, evenLually requlrlng more sesslons Lo achleve Lhe
deslred volume and shape.

19

rote|n fo|d|ng stress |n neurodegenerat|ve d|sease: an |nterp|ay between Lk stress
and autophagy
Claudlo PeLz

8lomedlcal neurosclence lnsLlLuLe (8nl), laculLy of Medlclne, unlverslLy of Chlle &
CenLer for Molecular SLudles of Lhe Cell, lC8M, unlverslLy of Chlle, Chlle.

A8S1kAC1: 1he mosL common neurodegeneraLlve dlseases, such as Alzhelmer's
ulsease, arklnson's ulsease, AmyoLrophlclaLeral sclerosls, and PunLlngLon's dlsease,
affecL mllllons of people worldwlde, buL Lhere ls nelLher prevenLlve nor curaLlve Lherapy
for Lhem. 1hese dlseases share a common neuropaLhology, prlmarlly feaLurlng Lhe
presence of abnormal proLeln lncluslons conLalnlng speclflc mlsfolded proLelns. 8ecenL
evldence lndlcaLes LhaL alLeraLlon ln organelle funcLlon ls a common paLhologlcal
feaLure of proLeln mlsfoldlng dlsorders. 1he endoplasmlcreLlculum (L8) ls an essenLlal
comparLmenL for proLeln foldlng, maLuraLlon, and secreLlon. Slgns of L8 sLress have
been exLenslvely descrlbed ln mosL experlmenLal models ofneurologlcal dlsorders. L8
sLress ls caused by funcLlonal dlsLurbances, whlch resulL ln Lhe accumulaLlon of
unfolded/mlsfolded proLelns aL Lhe L8 lumen. 1o cope wlLhL8 sLress, cells acLlvaLe an
lnLegraLed slgnallng response Lermed Lhe unfolded roLeln 8esponse (u8), whlch alms
Lo reesLabllsh homeosLasls Lhrough LranscrlpLlonal up regulaLlon of genes lnvolved ln
proLeln foldlng, quallLyconLrol and degradaLlon paLhways. ln Lhls Lalk we overvlew our
efforLs Lo assess Lhe role of L8 sLress ln proLeln mlsfoldlng dlsorders, and dlscuss
posslble sLraLegles Lo LargeL Lhe u8 wlLh LherapeuLlc beneflLs.

SupporL from lCnuLC?1 no. 1100176, lCnuA granL no. 13010006, Mlllennlum
lnsLlLuLe no. 09-013-l, Muscular uysLrophy AssoclaLlon, Alzhelmer AssoclaLlon, , and
Lhe Mlchael !. lox loundaLlon for arklnson research (Lo CP)



20

Meta| stress on p|tu|tary factors |n Cypr|nuscarp|o

kausel, C., valenzuela, C.L., erez, A.S.A., Munoz, A., SLolzenbach, l., 8omero, A.,
llgueroa, !.

lnsLlLuLo de 8loqulmlca y Mlcroblologla, laculLad de Clenclas, unlversldad AusLral de
Chlle, valdlvla.

lacLors of Lhe hypoLhalamo-hypophyseal axls consLlLuLe Lop hlerarchlcal regulaLors Lo
adapL an organlsm Lo changlng envlronmenLs.
WlLh Lhe alm Lo develop blomarkers for changes ln Lhe aquaLlc envlronmenL we
characLerlze blologlcally relevanL meLal effecLs ln Lhe conLexL of whole organlsm.
1herefore we evaluaLed gene expresslon of a serles of facLors ln adulL cyptlooscotplo.
LffecL of meLal was deLermlned ln Llssues from carp ln[ecLed lnLra perlLoneally wlLh
ZnCl
2
(0,6mg/kg) for Lhree consecuLlve days and analysed on Lhe fourLh day by 81-qC8.
Clearly lncrease of classlcal marker gene meLalloLhlonelnln llver of meLal LreaLed respecL
Lo conLrol carp conflrmed physlologlcal response and valldaLed dlfferenLlal effecL on
expresslon of a serles of hormones and LranscrlpLlon facLors. 1he meLal response
LranscrlpLlon facLor M1l1 was lmmuno deLecLed ln a subseL of somaLoLrophs ln
plLulLarygland and mlghL play a role ln decrease of CP expresslon ln Zn LreaLed respecL
Lo conLrol carp. We are currenLly an alyslngeplgeneLlc markers Lo complemenL analyses
of dynamlc envlronmenL - gene lnLeracLlones.

21

nomo muscu|us, more than [ust a mouse w|th human b|ood?
Maroun khoury
unlversldad de Los Andes, Chlle.
Pumanlzed" mlce are a promlslng LranslaLlonal model for sLudylng human
hemaLopolesls and relaLed dlseases. 1he Lrlple Lransgenlc mouse sLraln (nCu-sclJ
ll2ty
ooll
) LhaL lack adapLlve lmmune funcLlon, supporL helghLened levels of human
hemaLolymphold engrafLmenL. Pumanlzed mlce can supporL sLudles noL only ln areas of
lmmunology, buL also ln LransplanLaLlon, lnfecLlous dlseases, and cancer. 1hese models
are parLlcularly valuable ln experlmenLaLlon where Lhere ls no approprlaLe small anlmal
model of Lhe human dlsease, as ln Lhe case of cerLaln vlral lnfecLlons. 1hls presenLaLlon
wlll deLalls Lhe creaLlon of humanlzed mlce by engrafLmenL of lmmunodeflclenL mlce
wlLh hemaLopoleLlc sLem cells derlved from human umblllcal cord blood and oLher
Llssues, and dlscusses conslderaLlons for Lhe usage of Lhe model for speclflc appllcaLlons.

22

Sp|na| Cord kegenerat|on |n kenopus
!uan Larraln
CenLer for Aglng and 8egeneraLlon, Mlllenlum nucleus ln 8egeneraLlve 8lology,
onLlflcla unlversldad CaLllca de Chlle, Chlle.




23

now to repa|r a broken heart?
Serglo Lavandero
Center for Molecular Studies of the Cell, Faculty of Chemical and Pharmaceutical
Sciences/ Faculty of Medicine, University of Chile, Santiago, Chile.
24

C||n|ca| cha||enges and research opportun|t|es |n |ast|c Surgery
Pans-CunLher Machens

ueparLmenL of lasLlc Surgery and Pand Surgery, kllnlkum rechLs der lsar, 1echnlsche
unlverslLaL Munchen, Munlch, Cermany.


1he cllnlcal fleld of lasLlc Surgery has Loo long and generally been mlsundersLood as a
playground for beauLy surgeons only. ln facL, lasLlc Surgery comprlses far more LhaL
blunL AesLheLlcs, buL offers an armamenLarlum of dlfferenL Lechnlques Lo help paLlenLs,
who are dlsabled due Lo a lack of cells or Llssues for heredlLary, posLLraumaLlc,
posLoncologlc or oLher dlsease-relaLed reasons. lurLhermore, lasLlc Surgery ls sLrongly
relaLed Lo Lhe whole fleld of 8egeneraLlve and 8econsLrucLlve Medlclne lncludlng 1lssue
Lnglneerlng, whlch fuels Lhe sclenLlflc communlLy slnce more Lhan a decade now wlLh
new and groundbreaklng lnslghLs lnLo funcLlonal human blology, e.g. sLem cell research.
1hls presenLaLlon glves lnslghL lnLo Lhe acLual work of lasLlc Surgeons, Lhe cllnlcal
challenges, we are faclng and descrlbes relevanL experlmenLal models. lLs alm ls Lo polnL
ouL research opporLunlLles, Lo flnd new soluLlons for unsolved cllnlcal problems and Lo
fosLer collaboraLlons wlLh LhemaLlcally relaLed sclenLlflc flelds llke blology, maLerlal and
compuLer sclences.


23

Lp|genet|c contro| of express|on of bone-master regu|ators dur|ng osteob|ast ||neage
comm|tment

MarLln MonLeclno, Adrlana 8o[as, Pugo Sepulveda, 8odrlgo Agullar, 1aLlana Cpazo.

CenLer for 8lomedlcal 8esearch and lCnuA CenLer for Cenome 8egulaLlon, Andres
8ello unlverslLy, SanLlago, Chlle.

8unx2 and CsLerlx (Csx) LranscrlpLlon facLors are essenLlal for bone developmenL and
osLeoblasL dlfferenLlaLlon as Lhey conLrol Lhe expresslon of a slgnlflcanL number of
down-sLream bone-phenoLyplc LargeL genes. Pere, we presenL evldence
supporLlngeplgeneLlc conLrol ofLranscrlpLlonal acLlvaLlon of 8unx2 and Csx genes durlng
commlLmenL ofmesenchymal and osLeoprogenlLor cells Lo Lhe osLeoblasLlc llneage.
AddlLlonally, we ldenLlfy key eplgeneLlc mechanlsms LhaL conLrlbuLe Lo sllenclng of Lhese
Lwo bone-masLer regulaLors durlng dlfferenLlaLlon of mesenchymal cells Lo non-
osLeoblasLlc cells. Cur resulLs demonsLraLe LhaL LranscrlpLlon of 8unx2 and Csx genes ln
osLeoprogenlLor cells lnvolves chromaLln remodellng processes LhaL are assoclaLed wlLh
Lhe presence of speclflc paLLerns of posL-LranslaLlonal modlflcaLlons ln hlsLones P3 and
P4 aL Lhe promoLer reglon of boLh genes. lmporLanLly, we demonsLraLe LhaL key
members of Lhe polycomb and LrlLhorax group of proLelns are necessary durlng 8M2-
dependenL LranscrlpLlonal acLlvaLlon of Lhe 8unx2 and Csx genes, lndlcaLlng LhaL Lhey
are relevanL componenLs durlng Lhe esLabllshmenL and malnLenance of Lhese eplgeneLlc
paLLerns.





26

1he ro|e of ag|ng |n ad|pose-der|ved stem ce||s and the consequences for the|r |n-v|tro
and |n-v|vo use

xueylng ?an and Andreas nussler

8C 1rauma PosplLal 1ublngen, Lberhard karls unlverslLy 1ublngen, 1ublngen, Cermany.

1here ls lncreaslng evldence LhaL mesenchymal sLem cells can dlfferenLlaLe lnLo hepaLocyLe-
llke cells (PCLs). Powever many open quesLlons remaln ln order Lo use Lhese cells rouLlnously
ln drug developmenL. Ad-MSCs were lsolaLed from abdomlnal adlpose Llssue of young (<40a)
and old (>60a) donors accordlng Lo eLhlcal guldellnes. rollferaLlon (Alamar blue assay, kl67
SLalnlng) and expresslon of plurlpoLency markers (CcL4, nanog, Sox2, c-Myc, kLl4) were
deLermlned. PepaLlc dlfferenLlaLlon wlLh/wlLhouL 3-azacyLldlne (AZA) and 8lx was lnlLlaLed ln
passage 3, afLerwards enzymaLlc and meLabollc acLlvlLles of PLCs were measured. Ad-MSCs
derlved from old donors demonsLraLed less prollferaLlon capaclLy Lhan young ones. 1he global
meLhylaLlon sLaLus ln young Ad-MSCs was lower Lhan ln old ones, whlch concurred wlLh Lhe
expresslon of nanog and Sox2 belng sLronger ln younger donors. Ad-MSC-derlved PLCs from
young and old donors pre-LreaLed wlLh AZA or 8lx, demonsLraLed slmllar enzymaLlc and
meLabollc acLlvlLles, ln some cases Lhey reached slmllar levels as seen ln human hepaLocyLes.
Ad-MSCs from old donors wlLhouL AZA or 8lx pre-LreaLmenL showed a much lower hepaLlc
dlfferenLlaLlon capaclLy Lhan young ones. 8educed expresslon of nanog and Sox2 may be a
causaLlve facLor for poor prollferaLlon of old Ad-MSCs and Lhelr consequenLly lower meLabollc
capaclLy afLer hepaLlc dlfferenLlaLlon. lurLhermore, we wlll dlscuss Lhe effecLs of an ln-vlvo
appllcaLlon.
27

1he ro|e of the ||ver |n bone damage: 1wo mode|s to study hepat|c osteodystrophy

Andreas nussler and Sabrlna LhnerL

8C 1rauma PosplLal 1ublngen, Lberhard karls unlverslLy 1ublngen, 1ublngen, Cermany.

8ackground: aLlenLs wlLh chronlc llver dlsease frequenLly suffer from alLeraLlons ln
bone mlneral meLabollsm resulLlng ln a decreased bone mlneral denslLy (8Mu), malnly
osLeoporosls or osLeopenla, deflned as hepaLlc osLeodysLrophy (PCu). PCu ls a
mulLlfacLorlal dlsease whose regulaLory mechanlsms are scarcely lnvesLlgaLed. lor
beLLer undersLandlng of Lhe underlylng mechanlsms we here developed a mouse model
LhaL represenLs Lhe human slLuaLlon and can easlly be Lransferred Lo oLher mouse
sLralns, e.g. gene knock-ouL or over-expresslng mlce, ln order Lo lnvesLlgaLe speclflc
gene funcLlon.
Methods and I|nd|ngs: We lnduced llver damage ln C378l6/n mlce by repeLlLlve
ln[ecLlons of CCl
4
(n=9). AfLer 6 weeks Lhese mlce develop severe llver flbrosls (P&L and
Slrlus 8ed sLalnlng) comparable Lo llver flbrosls ln paLlenLs (n>23). uurlng Lhls perlod
changes ln bone sLrucLure evolved (C1 analysls). CCl
4
LreaLed mlce have slgnlflcanLly
decreased 8Mu (297.331.1 mg PA/cm
3
vs. mg 211.616.7 PA/cm
3
, p<0.03), bone
volume (1.100.08 mm3 vs. 0.610.18 mm
3
, p<0.01), Lrabecular number (3.180.24 vs.
4.070.31, p<0.03) and Lhlckness (0.0690.003 mm vs. 0.0330.002 mm, p<0.01). ln
PCu paLlenLs alLered vlLamln u meLabollsm resulLs ln decreased 23-CP-vlLamln u serum
levels (133.323.4 nmol/l vs. 37.92.7 nm/l, p<0.001). Comparable Lo PCu paLlenLs, CCl
4

LreaLed mlce show decreased C?27A1, C?281, vlLamln u blndlng proLeln CC and
lncreased uPC87 hepaLlc gene expresslon. lurLhermore, boLh PCu paLlenLs and CCl
4

LreaLed mlce produce excesslve amounLs of acLlve 1Cl- (~3.3-fold, p<0.001) and whlch
has been shown Lo lnhlblL osLeoblasLs funcLlon lo vltto.
Conc|us|ons: 1he here presenLed mouse model recaplLulaLes several regulaLory key
mechanlsms of hepaLlc osLeodysLrophy and suggesLs a key role of 1Cl- ln llver-bone
communlcaLlon. lurLhermore, Lhe severlLy of llver dlsease ls closely documenLed and
slgnlflcanL changes on bone sLrucLure and mlnerallzaLlon can be observed by C1
analysls already afLer 6 weeks LreaLmenL wlLh CCl
4
.


28

Stem Ce||s and Ad|pogenes|s - |n v|tro and |n v|vo 1|ssue Lng|neer|ng
norberL allua
ueparLmenL of lasLlc and 8econsLrucLlve Surgery, Pand Surgery - 8urn CenLer,
unlverslLy PosplLal of Lhe 8W1P, Aachen, Cermany.
Adlpose Llssue ls a blologlcal and dynamlc flller wlLh a volumeLrlc and a regeneraLlve
effecL wldely used ln lasLlc and 8econsLrucLlve Surgery for Lhe fllllng of volume deflclLs
and conLour deformlLles. AparL from Lhe mere volume effecLs, faL Lransfer has also
re[uvenaLlon effecLs resulLlng ln Lhe reducLlon of wrlnkles by enhanclng dermal collagen
producLlon. ln Lhls conLexL, posslble reasons such as Lhe presence of adulL mesenchymal
sLem cells, Lhe so called adlpose-derlved sLem cells (AuSC) whlch occur abundanLly ln
adlpose Llssue and relevanL growLh facLors (e.g. blCl, lCl, uCl, vLCl) ln Lhe
Lransferred faL Llssue wlll be dlscussed.
lurLhermore, an overvlew wlll be presenLed abouL Lhe lsolaLlon of AuSCs and meLhods
of Lhelr dlfferenLlaLlon. lacLors, such as llposucLlon procedures, Lhe lsolaLlon Lechnlques,
purlflcaLlon sLeps and cell culLure condlLlons affecLlng Lhe yleld and behavlour of AuSCs
are dlscussed. AddlLlonally, Llssue englneerlng approaches of sofL Llssue wlll be
presenLed and Lhe lmporLance and poslLlve effecLs of adlpose-derlved growLh facLors on
faL grafL survlval and Llssue englneerlng wlll be dlscussed.
ln concluslon, adlpose Llssue and AuSCs are valuable maLerlal for ongolng and fuLure
sLem cell based Lheraples wlLh a wlde range of posslble cllnlcal appllcaLlon. Powever,
aLLenLlon musL be pald regardlng poslLlve and posslble negaLlve slde effecLs of adlpose
derlved growLh facLors and cyLoklne uLlllzaLlon.

29

Neogen|n- 1 modu|ates ang|ogenes|s |nduced by netr|n-1 produced by human
wharton's [e||y mesenchyma| stem ce||s (WI-MSC)
Carollna CrLlz, CynLhla vlllarroel, CaLallna rleLo & vernlca alma
LaboraLory of SLem Cells and uevelopmenL, laculLy of Sclences, unlverslLy of Chlle,
Chlle.

Introduct|on: Crlglnally ldenLlfled as axonal guldance molecules, neLrlns are lamlnln-llke
secreLed proLelns LhaL have been recenLly lnvolved ln anglogenesls and blood vessels
neLwork formaLlon, dependlng on Lhelr physlologlcal conLexL. A cruclal LargeL recepLor
for nCs durlng embryonlc developmenLls Lhe neogenln-1 recepLor, however lLs role
durlng an glogenesls ls unknown. 1he alm of Lhls work was Lo elucldaLe Lhe role of W!-
MSC produced neLrlns ln Lhe lnducLlon of anglogenesls acLlng Lhrough Lhelr puLaLlve
recepLor neogenln-1, presenL ln human umblllcal chord endoLhellal cells (PuvLC).
Methodo|ogy: We evaluaLed neLrln-1 and 4 ln W!-MSC and neogenln-1 ln PuvLC cell
culLures Lhrough 81-C8, W8, lACS and lmmunohlsLochemlsLy ln umblllcal cord
secLlons. Anglogenlc ablllLy of condlLloned medlum, obLalned from W!-MSC was assayed
wlLh scraLch and MaLrlgelLubule formaLlon assays. 1o sLudy Lhe relaLlve conLrlbuLlon of
Lhe neogenln-1 recepLor we performed experlmenLs uslng PuvLC Lransduced wlLh
lenLlvlral vecLors expresslng shneo-1.1o ascerLaln whlch neLrln llgand medlaLed
anglogenlc acLlvlLy by blndlng Lo neogenln-1 we made use ofshneLrln1and/or shneLrln4
vecLors.
kesu|ts: PuvLC express neogenln-1, whlle W!-MSC release neLrln-1 and 4. We
demonsLraLe LhaL W!-MSC condlLloned medlum promoLes Lubule formaLlon and
mlgraLlon of PuvLC.
D|scuss|on: We propose a cross-Lalk beLween neLrlns, secreLed by W!-MSC, and
neogenln-1 presenL ln PuvLC. 1hls communlcaLlon may have deep lmpllcaLlons ln Lhe
modulaLlon of several processes such as anglogenesls and could be useful for opLlmlzlng
LherapeuLlc approaches.
Iund|ng: lCnuA 13090007 and lCnuLl u09l1047.

30

1oward a manufactur|ng process of human ad|pose stroma| ce||s for ce|| therapy:
pre||m|nary resu|ts

karaglannl Marlanna
1
, 8leback karen
1
, kluLer Parald
1
, lrlnger Marlon
2
, Wlgand Sylvla
2
,
Machens Pans-CunLher
2
, apadopulos nlkolaos
2

1
lnsLlLuLe of 1ransfuslon Medlclne and lmmunology, Medlcal laculLy Mannhelm,
Peldelberg unlverslLy, Cerman 8ed Cross 8lood uonor Servlce 8aden-WurLLemberg -
Pessen.
2
ueparLmenL of lasLlc Surgery & Pand Surgery, unlverslLy PosplLal rechLs der
lsar, Munlch 1echnlcal unlverslLy, Cermany.

Introduct|on: Adlpose sLromal cells (ASC) can be lsolaLed ln a hlgh frequency from
adlpose Llssue by low rlsk llposucLlon. 1hey show mulLlllneage dlfferenLlaLlon poLenLlal,
lmmune modulaLory and pro-regeneraLlve capaclLles. 1hus, Lhere ls a rapldly growlng
lnLeresL for ASC as candldaLes for cllnlcal appllcaLlon ln cell Lherapy.
1he ldea of sLem cell Lherapy sounds slmple, however cell Lherapy requlres a complex,
mulLlsLep process Lo LranslaLe research flndlngs lnLo cllnlcal rouLlne, cell LherapeuLlcs
have Lo comply wlLh regulaLory guldellnes, be safe and efflclenL. Accordlngly, ln order Lo
esLabllsh cllnlcal proLocols for ASC, lL ls lmporLanL Lo undersLand and carefully conLrol
Lhe producLlon process and Lo deflne measures LhaL rellably predlcL safeLy and efflcacy
of cell LherapeuLlcs.

Mater|a| and Methods: WlLhln a flrsL sLep ln esLabllshlng manufacLurlng proLocols for
ASC, we asked wheLher Lhe procuremenL meLhod and anaLomlcal locallsaLlon of adlpose
Llssue affecL ASC quallLles. 1herefore, llpoasplraLes were obLalned uslng Lwo sLerlle
devlces and derlved from dlfferenL anaLomlcal slLes, lncludlng abdomlnal and Lhlgh
Llssues. LlpoasplraLes were processed Lo lsolaLe Lhe sLromal vascular fracLlon. 1hese
were analysed accordlng Lo Lhe cell yleld/g Llssue, cell vlLallLy, precursor frequency,
lmmune phenoLype and expanded Lo derlve ASC. ASC were characLerlsed by flow
cyLomeLry and dlfferenLlaLlon Lowards Lhe adlpogenlc and osLeogenlc llneage.

kesu|ts: AfLer evaluaLlng Lhe flrsL 44 speclmens from 28 paLlenLs we show a Lendency
for hlgher number of vlLal cells/g ln male paLlenLs wlLh a 8Ml over 23 when uslng 11l
devlce (1rans1lssuelllLron 300, ShlpperL Medlcal 1echnologles Corp., uSA). 1he donor-
slLes were all comparable, alLhough more vlLal cells/g were presenL when faL was Laken
paraverLebral, alLhough only ln 2 speclmens!

Conc|us|on: vla Lhls analysls we alm Lo esLabllsh a cllnlcal proLocol for ASC lsolaLlon and
Lo move Lo a well deflned cllnlcal sLudy, focuslng on sLandardlsaLlon concernlng Lhe
applled cell LherapeuLlc fracLlon.

31

|acenta| stem ce||, from bas|c research to c||n|ca| app||cat|ons

1,2
San MarLln S.,
3
venegas L.,
2
Moller A.,
1,2
vlllena !.,
1,2
rraga M

1
8lomedlcal 8esearch CenLre, School of Medlclne, unlversldad de valparalso, Chlle.
2
lnblocrloLec.
3
CphLhalmology unlL, van 8uren PosplLal, valparalso, Chlle.

CesLaLlonal Llssue such as Lhe placenLa, amnloLlc membranes, and amnloLlc fluld are
usually dlscarded followlng blrLh, and lLs use does noL ellclL eLhlcal debaLe. Moreover,
several llne research showed LhaL cells from Lhls Llssue are a good cholce for
regeneraLlve medlclne because of boLh Lhelr phenoLyplc plasLlclLy and
lmmunomodulaLory capablllLy. 1he amnlon ln parLlcular, ls a Lhln, avascular membrane
derlved from embryo, whlch ls composed of an eplLhellal layer and an ouLer layer of
connecLlve Llssue. ls lL readlly avallable and easlly procured wlLhouL lnvaslve procedures.
uslng amnloLlc membrane grafLs has been an effecLlve meLhodology ln eye surface
paLhologles such as bullous keraLopaLhy. 1hls dlsease ls characLerlzed by an endoLhellal
pump dlsorder whlch generaLes a chronlc corneal edema evolvlng Lo Lhe producLlon of
veslcles and bullae whlch produce chronlc eye paln and vlsual aculLy decrease.
ueflnlLlve LreaLmenL for Lhose paLlenLs sufferlng from bullous keraLopaLhy (8k) ls
corneal LransplanL. Powever, donaLlon ls noL always avallable and Lhus requlres long
walLlng Llmes. As a resulL of Lhls delay, paLlenLs have Lo be managed palllaLlvely wlLh
conLacL lenses LhaL prevenL Lhe corneal eplLhellum from falllng. uslng conLacL lenses ln
Lhls slLuaLlon ls assoclaLed Lo compllcaLlons such as: corneal neovascularlzaLlon, lens
dlsplacemenL or loss, lnfecLlons, allLogeLher wlLh Lhe economlc expense assoclaLed Lo
Lhe use of lenses.
Cur group compares Lhe use of amnloLlc membrane grafLs versus conLacL lenses ln
paLlenLs sufferlng from 8k awalLlng a corneal LransplanL wlLh respecL Lo: ocular paln
rellef, eplLhellal defecLs and corneal bullae, vlsual aculLy and corneal neovascularlzaLlon.
1o answer Lhls ob[ecLlve, a conLrolled cllnlcal assay was performed wlLh paLlenLs
supporLlng a cllnlcal dlagnosls of 8k awalLlng a corneal LransplanL. aLlenLs had been
dlagnosed beLween 2008 and 2011 and all came from Lhe CphLhalmology unlL of Lhe
PosplLal Carlos van 8uren ln valparalso, Chlle. aLlenLs were randomlzed lnLo 2 groups:
amnloLlc membrane and LherapeuLlc conLacL lenses. Lye paln lnLenslLy (Snelle
quesLloner), bullae and corneal eplLhellal defecLs presence (Analog vlsual scale), corneal
neovascularlzaLlon, vlsual aculLy and compllcaLlons (blomlcroscopy) were compared
among Lhe Lwo groups menLloned, for a perlod of 6 monLhs.
uurlng Lhe observaLlon perlod, 20 paLlenLs wlLh 8k awalLlng corneal LransplanL were
sLudled. aln from Lhe surgery was sLaLlsLlcally slgnlflcanL lower aL 7 (p=0.003) and 30
days (p=0.002) buL hlgher aL 90 days (p=0.042). SLaLlsLlcally slgnlflcanL dlfferences
concernlng bullae / eplLhellal defecLs presence, corneal neovascularlzaLlon and vlsual
aculLy were noL observed, beLween boLh groups.
AmnloLlc membrane LransplanL consLlLuLes an efflclenL and safe alLernaLlve Lo Lhe use of
LherapeuLlc conLacL lenses Lo rellef ocular paln aL shorL Lerm ln paLlenLs wlLh 8k.

Acknow|edgment: Cl 03/2006 (unlversldad de valparalso, Chlle) and CCnlC?1 (AC1-73),
Chlle.
32







A8S1kAC1S CS1Lk kLSLN1A1ICNS
(A|phabet|c order)

33

Lp|genet|c s||enc|ng of key bone phenotyp|c genes dur|ng neurona| deve|opment
8. Agullar
1
, l. 8usLos
1,2
, 8. Penrlquez
1
, A. 8o[as
1,3
, 8. van ZunderL
1
and M. MonLeclno
1
1
CenLer for 8lomedlcal 8esearch, Andres 8ello unlverslLy, SanLlago, Chlle,
2
unlverslLy of
Concepclon, Concepclon, Chlle,
3
unlverslLy of Chlle, SanLlago, Chlle.

8unx2 and CsLerlx LranscrlpLlon facLors are essenLlal for bone developmenL and
osLeoblasLlc dlfferenLlaLlon as Lhey regulaLe Lhe expresslon of key bone llneage-speclflc
genes. ln osLeoblasLs, eplgeneLlc mechanlsms LhaL lead Lo LranscrlpLlonal acLlvaLlon of
Lhese genes have been descrlbed, buL Lhelr sllenclng ln non-osLeoblasLlc cells remalns Lo
be elucldaLed. ln neurons, Lhe regulaLory neLworks LhaL may Lrlgger LranscrlpLlon of
osLeoblasL-speclflc genes are acLlve, alLhough are unable Lo promoLe bone-relaLed gene
expresslon. We alm Lo evaluaLe Lhe conLrlbuLlon of eplgeneLlc mechanlsms durlng
sllenclng of osLeoblasL-speclflc genes ln neuronal cells. Plppocampal Llssue from L18,
10, 30 and 90 raLs was obLalned Lo perform ChromaLln lmmunopreclplLaclon (Chl)
sLudles uslng anLlbodles dlrecLed agalnsL covalenL hlsLone modlflcaLlons and chromaLln
modlfylng enzymes. lL was found LhaL 8unx2 and CsLerlx promoLer sequences exhlblL
enrlchmenL of Lhe represslve P3k27me3 eplgeneLlc mark ln all Lhe sLages analyzed.
AddlLlonally, we deLecL Lhe presence of olycomb-group proLelns Lzh1 and Lzh2,
blndlng ln a developmenLal sLage-dependenL manner aL 8unx2 and CsLerlx promoLers,
correlaLlng Lhelr expresslon paLLerns. We also flnd Lhe heLerochromaLln-assoclaLed
P3k9me3 eplgeneLlc mark ln Lhe adulL sLage, LlghLly correlaLed wlLh decreased hlsLone
P3 aceLylaLlon and enrlchmenL of PlsLone ueaceLylases PuAC1 and PuAC2. llnally, we
flnd a sLrong unA meLhylaLlon paLLern aL bone-speclflc genes (e.g. CsLeocalcln) LhaL are
expressed aL laLe sLages durlng bone formaLlon. 1ogeLher, our resulLs supporL a model
where eplgeneLlc posL-LranslaLlonal modlflcaLlons ensure Lhe sllenclng of non-neuronal
genes durlng neuronal dlfferenLlaLlon.

SupporLed by CCnlC?1 24120931, unA8-ul-70-12/l, lCnuA 13090007, lCnuLC?1
1093073, lCnuLC?1 3110138, and lCnuLC?1 1101012.


34

Lffects of exposure to c|garette smoke on t|ssue regenerat|on |n zebraf|sh
Alvarez, Mar[orle
1
, SoLo, Claudlo
1
, Lgana, !ose 1oms
1,2
and Allende, Mlguel L.
1
1
lCnuA CenLer forCenome8egulaLlon, laculLad de Clenclas, unlversldad de Chlle,
2
ueparLmenL of lasLlc Surgery and Pand Surgery, kllnlkum rechLs der lsar, 1echnlsche
unlverslLaL Munchen, Cermany.

CxldaLlon ls esenclal for llfe, buL, ln excess, lL can lead Lo oxldaLlve sLress, wlLh Loxlc
effecLs on dlfferenL blologlcal sysLems. We seek Lo generaLe an anlmal model of
oxldaLlve sLress lnducLlon and llnk oxldaLlve sLaLus wlLh regeneraLlve capaclLy. We use
larval and adulL zebraflsh of dlverse Lransgenlc sLralns Lo monlLor cell survlval and
behavlor ln Lhe conLexL of regeneraLlon and evaluaLe Lhe effecL of oxldaLlve sLress on
Lhls process. lL ls known LhaL clgareLLe smoke ls hlghly oxldlzlng, LhaL lL can alLer
anglogenesls and Lhe regeneraLlve poLenclal ln mammals, buL Lhere are no good models
Lo assess Lhe mechanlsms by whlch Lhese changes are generaLed. lurLhermore, Lhere
are no efflclenL ways Lo evaluaLe Lhe effecLs of drugs or LoxlcanL removal sysLems of pro-
oxldanLs LhaL may decrease Lhe effecLs of clgareLLe smoke. Pere, we have evaluaLed Lhe
effecLs of clgareLLe smoke on regeneraLlon of Lhe Lall fln. We flnd LhaL LreaLed flsh
regeneraLe poorly compared Lo conLrols, posslbly due Lo poor vascularlzaLlon. ln
addlLlon, we have observed a poLenL effecL on Lhe lnflammaLory response lnduced by
Lall fln secLlonlng ln conLrols vs. smoke exposed anlmals. llnally, we have evaluaLed Lhe
regeneraLlve capaclLy of aged flsh compared Lo young flsh ln order Lo deLermlne
wheLher aglng lmpacLs Lhe ablllLy of Llssues Lo regeneraLe, besldes we evaluaLed Lhe
anLloxldanL effecL of ttoostesvetottol on larval zebraflsh. Cur sLraLegy wlll lmprove our
ablllLy Lo evaluaLe Lhe effecLs of oxldaLlve sLress as well as Lo measure Lhe anLloxldanL
acLlvlLy of dlfferenL molecules ln vlvo uslng Lhe zebraflsh as a bloassay sysLem.

33

1rypanosama cruz| Ca|ret|cu||n, a paras|te prote|n that |nh|b|ts bone regenerat|on

!. lgnaclo Arlas
1
, aLrlcla 8ravo
1
, ChrlsLopher PamllLon-WesL
1
, lsmael Maldonado
2
and
ArLuro lerrelra
2
1
uepLo. Clenclas Cllnlcas, laculLad de Clenclas veLerlnarlas, unlversldad de Chlle,
SanLlago, Chlle,
2
lC8M, laculLad de Medlclna, unlversldad de Chlle, SanLlago, Chlle.

ln orLhopaedlcs, Lhe use of facLors LhaL enhance granulaLlon Llssue formaLlon and
prevenL or delay new bone regeneraLlon ls someLlmes deslrable. CalreLlculln (C81), a
unlque endoplasmlc reLlculum lumlnal Ca2+-blndlng chaperone wldely dlsLrlbuLed ln
eukaryoLlc cells, ls lnvolved ln many cellular funcLlons. Among Lhem, C81 has an
lmporLanL lnfluence ln cuLaneous wound heallng and dlverse processes assoclaLed wlLh
cuLaneous repalr, lnhlblLlon of anglogenesls, promoLlon of cell adheslon and anLlLumor
effecL. Cne of Lhe molecules lnvolved ln several aspecLs of Lhe hosL-paraslLe lnLerplay ls
1rypanosoma cruzl calreLlculln (1cC81), whlch ls hlghly homologous Lo human
calreLlculln (PuC81). Pere, recomblnanL (r)PuC81 and r1cC81 are compared on Lhelr
ablllLles Lo affecL bone heallng ln a rabblL model. ln molar Lerms, r1cC81 ls more efflclenL
Lhan rPuC81 ln decreaslng of bone gap brldglng ln rabblL unlcorLlcal bone osLeoLomles.




36

hotosynthet|c Derma| Scaffo|d for Sk|n kegenerat|on
Chvez Mn
1
, Schenck 1L
1
, Popfner u
1
, ClunLa 8L
1,2
, Machens PC
1
, 8ohne Av
3
, nlckelsen
!
3
and Lgana !1
1,4
.

1
ueparLmenL of lasLlc Surgery and Pand Surgery, unlverslLy PosplLal rechLs der lsar,
1echnlsche unlverslLaL Munchen, Cermany,
2
Pand, lasLlc and AesLheLlc Surgery,
kllnlkum der Ludwlg MaxlmlllansunlverslLaL, Munchen, Cermany,
3
Molekulare
flanzenwlssenschafLen, 8lozenLrum Ludwlg-Maxlmlllans-unlverslLaLMunchen, lanegg-
MarLlnsrled, Cermany,
4
lCnuA CenLer for Cenome 8egulaLlon, laculLad de Clenclas,
unlversldad de Chlle, SanLlago, Chlle.

1lssue englneerlng has opened a new LherapeuLlc avenue LhaL promlses a revoluLlon ln
regeneraLlve medlclne. Powever, Lo daLe Lhe regeneraLlve poLenLlal of englneered
Llssues ls llmlLed and Lhe cllnlcal resulLs are dlsappolnLlng. 1hls ls malnly aLLrlbuLed Lo
Lhe unsolved problem of poor vascularlzaLlon and Lhe resulLlng hypoxla of englneered
Llssues. Pere we propose an alLernaLlve source of oxygen Lo blood vessel-perfuslon by
uslng phoLosynLheLlc scaffolds. We demonsLraLe LhaL Lhe unlcellular and phoLosynLheLlc
alga ChlamydomonasrelnhardLll can be culLured ln scaffolds for dermal regeneraLlon
showlng hlgh blocompaLlblllLy and phoLosynLheLlc acLlvlLy. Moreover, Chlamydomonas
can be co-culLured wlLh flbroblasLs, decreaslng Lhe hypoxlc response under low oxygen
culLure condlLlons. LngrafLmenL of phoLosynLheLlc scaffolds ln a full skln defecL ln nude
mlce generaLes symbloLlc Llssues composed of algae and murlne cells. 1he resulLs
obLalned here represenL Lhe flrsL sLep for englneerlng human auLoLrophlc Llssues.

37

Ce||u|ar character|zat|on of the response to sp|na| cord |n[ury of "#$%&'()*#+,(!
Cabrlela Ldwards and !uan Larraln

CenLer for Aglng and 8egeneraLlon, Mlllenlum nucleus ln 8egeneraLlve 8lology,
onLlflcla unlversldad CaLllca de Chlle.

Splnal cord ln[ury (SCl) ln mammalsleads Lo haemorrhage and cell deaLh resulLlng ln
cavlLles formaLlon and ls followed by a secondary damage whlch lncludes an
lnflammaLory response, oedema,demyellnlzaLlonandasLrogllosls. All Lhls process
culmlnaLes wlLh Lhe formaLlon of a gllal scar, Lhe prlnclpal lmpedlmenL for axon
regeneraLlon. ln conLrasL, amphlblans such anuran larvae of \eooposare able Lo
regeneraLe afLer SCl, however, Lhls regeneraLlve capaclLy ls losL durlng meLamorphosls.
1he cellular mechanlsms responslble for Lhls process are unknown. Cur hypoLhesls ls
LhaL regeneraLlon of Lhe splnal cord ln pre-meLamorphlc sLages of \eooposloevls ls
allowed by apermlsslveasLrogllosls, whlch becomes lnhlblLory for regeneraLlondurlng
meLamorphosls".
Mater|a| and Methods: Cur sLudy model lssplnal cord LransecLlon ln[ury ln pre-
meLamorphlc sLages (sL30 and sL36) and posL-meLamorphlc sLage (sL66) of
\eooposloevls. 1he analysls was performed uslng cryosecLlons and paraffln secLlons of
splnal cord, hlsLologlcal sLalnlng andlmmunofluorescence. 1he LoLal dlsLance moved
afLer ln[ury was evaluaLed by Lhe An?maze program.
kesu|ts: PlsLologlcal analyslsshowedLhaLsL30\eooposparLlallyreorganlzedLhe splnal cord
Llssue afLer 10 days posL ln[ury, whlch correlaLes wlLh Lhe parclal recovery of Lhe LoLal
dlsLance moved, unllke sL36 and sL66 anlmals. lmmunefluorescence analysls showed a
dlfferenLlal expresslon of asLrogllosls markers such a ClA, vlmenLln and some
exLracellular maLrlx proLelns such as flbronecLln, collagen and chondrolLln sulfaLe
proLeoglycans.
D|scuss|on: 1hese dlfferencesmay accounL for Lhe regeneraLlve ablllLles of pre-
meLamorphlc sLages and Lhe lack of Lhere afLer meLamorphosls ln \eooposloevls.
Grants: Mln8L8 07/011, CenLer for Aglng and 8egeneraLlon CA8L.
38

S||enc|ng Dgcr8 and over-express|ng L|n28 |n mesenchyma| stem ce||s (MSC)
estab||shes a cr|t|ca| funct|on for m|crokNAs |n MSC
llerro lA, kalomolrls S, 8eegle ! and nolLa !A
lnsLlLuLe for 8egeneraLlve Cures, laculLy of Medlclne, unlverslLy of Callfornla aL uavls,
uSA.
MulLlple recenL sLudles have evaluaLed Lhe role of mlcro8nAs (ml8nAs)
ln mesenchymal/skeleLal sLem cells (MSC). Pere, we addressed Lwo baslc quesLlons
relaLed Lo Lhe funcLlon of ml8nAs ln MSC: (1) whaL ls Lhe consequence of sllenclng Lhe
enLlre ml8nA machlnery ln MSC and (2) whaL ls Lhe role of leL-7 ml8nAs, Lhe mosL hlghly
expressed famlly of ml8nAs ln MSC? Pence, we Lransduced human MSC wlLh Lwo
dlfferenL sh8nAs Lo suppress ugcr8, an essenLlal componenL for Lhe blogenesls of all
ml8nAs. ugcr8 reducLlon lnhlblLed cell prollferaLlon and sLrongly reduced adlpogenlc
and osLeogenlc dlfferenLlaLlon of MSC ln vlLro. ueep sequenclng analysls of small 8nAs
ln MSC revealed LhaL Lhese cells express hlgh levels of mulLlple leL-7 ml8nAs. As a
prellmlnary approach, we over-expressed Lhe plurlpoLency facLor Lln28, whlch ls a well
know lnhlblLor of leL-7. Cver-expresslon of Lln28 dld noL affecL prollferaLlon, buL
lnhlblLed boLh adlpogenesls and osLeogenesls of MSC ln vlLro, alLhough Lo a lesser
exLenL Lhan sllenclng ugcr8 dld. uurlng osLeogenesls, over-expresslon of Lln28 lnhlblLs
maLrlx maLuraLlon and sLages afLerwards, suggesLlng LhaL leL-7 only regulaLe laLe sLages
of osLeogenesls, whlle oLher ml8nAs musL regulaLe Lhe early phases. 1hese sLudles
deflne Lhe baslc role of ml8nAs and suggesL novel funcLlons Lo leL-7 ln MSC, whlch could
lmpacL Lhe LherapeuLlc use of MSC Lo LreaL crlLlcal bone defecLs.



39

Center for Sc|ence Commun|cat|on Un|vers|dad Andres 8e||o
naLalla Mackenzle
unlversldad Andres 8ello, vlcerrecLorla de lnvesLlgacln y uocLorado, SanLlago, Chlle.

1he new CenLer for Sclence CommunlcaLlon was creaLed Lo brlng unA8's research closer
Lo Lhe non-sclenLlflc communlLy and Lo help brldge Lhe exlsLlng gap beLween sclence
and socleLy Lhrough workshops, conferences, popular sclence arLlcles, sclence falrs and
oLher acLlvlLles. lL ls also responslble for lmprovlng publlc undersLandlng of sclence by
Lralnlng sclenLlsLs and [ournallsLs Lo communlcaLe sclence more effecLlvely. 1he CenLer
has recenLly won a governmenL granL Lo develop sclence workshops of bloLechnology,
nanoLechnology, aqulculLure, envlronmenLal sclence and sclence communlcaLlon for
[ournallsLs lnLeresLed ln coverlng sclenLlflc Loplcs ln publlc medla. AddlLlonally, Lhe
cenLer ls organlzlng a conference for CcLober 2013 on sclence communlcaLlon,
lnnovaLlon and pollLlcs, whlch wlll serve as a plaLform Lo encourage mulLldlsclpllnary
collaboraLlon beLween enLrepreneurs, sclenLlsLs, pollLlclans, communlcaLors and Lhe
communlLy. 1hese acLlvlLles, LogeLher wlLh our fuLure amblLlon Lo develop a medla lab
wlll Lake us closer Lo our maln goal: Lo fosLer lnnovaLlon based on sclence and reach ouL
Lo Lhe communlLy, uslng sclence as Lhe cornersLone for consLrucLlng a beLLer lnformed
socleLy.

40

Ietoscopy |n the pregnant rabb|t at m|d-gestat|on, a new mode| for |ntrauter|ne
endoscop|c p|ast|c surgery research?

apadopulos, nA
1,2
, ulmlLrascu, l
2
!" #$%&'()!" *+
2
, LeruL, 1L
2
, 8arkl, C
3
, Poehnke, C
1
,
8lemer, L
1
, & uepresL, !AM
2
1
uepL. of lasLlc Surgery, 1echnlcal unlverslLy Munlch, Cermany,
2
CenLre for Surglcal
1echnologles, kaLhollekeunlverslLelL Leuven, 8elglum,
3
karl SLorz Lndoskope, 1uLLllngen,
Cermany.

ln18CuuC1lCn: 8ecenLly Lhe lnLroducLlon of flberscopes and vldeo-endoscoplc
Lechnology has boosLed endoscopy. 1hls has renewed Lhe lnLeresL ln dlrecL vlsual access
Lo amnloLlc cavlLy for dlagnosLlc and LherapeuLlc procedures 1. 1oday feLoscopy ls used
Lo gulde LherapeuLlc procedures such as chorlonlc vessel coagulaLlon, umblllcal cord
llgaLlon and even feLal surgery 2. lL has been suggesLed LhaL Lhe endoscoplc approach
for Lhls purposes ls accompanled by lower maLernal morbldlLy and feLal morLallLy 3. 1he
alm of Lhls sLudy was Lo evaluaLe Lhe performance of dlrecL vlsuallzaLlon Lo Lhe feLus
and Lhe amnloLlc cavlLy ln a readlly avallable small anlmal model such Lhe mld-
gesLaLlonal rabblL model, uslng a convenLlonal vldeo camera and a 1.2mm flber-
endoscope, and Lo esLlmaLe lL's effecLs Lo Lhe feLus.
MA1L8lAL Anu ML1PCuS: 1welve Llme-daLed pregnanL new Zealand rabblLs were
housed Lwo days prlor Lo surgery. AL LwenLy-Lwo days gesLaLlonal age (CA, Lerm=32
days), Lhe feLoscopy was performed. AfLer premedlcaLlon anlmals were puL under
general face mask anaesLhesla uslng haloLhane (2-3). A llberal mldllne abdomlnal
lnclslon was made Lo exposeLhe pregnanL uLerus. MyomeLrlal lnclslon was done wlLh an
operaLlon mlcroscope (Carl Zelss!, magnlflcaLlon: 3-23x) and mlcrolnsLrumenLs 4.
AmnloLlc access was achleved Lhrough a 14-gauge needle puncLure, allowlng sallne
lrrlgaLlon and lnLroducLlon of a 10 cm long 1.2mm flberscope (karl SLorz Lndoscope)
connecLed Lo a slngle chlp CCu-camera, and meLalhallde llghL source. 1wo amnloLlc sacs,
one ln each uLerlne horn, randomly asslgned were used for feLoscopy (n=24). 1he
unLreaLed amnloLlc sacs were servlng as negaLlve conLrols (n=84). A 2-3mm myomeLrlal
lnclslon was made on Lhe avascular slde of Lhe uLerus, and Lhe chorlon was opened and
dlssecLed away from Lhe amnlon (llg 1). under mlcroscoplc conLrol Lhe 14-gauge needle
was lnserLed lnLo Lhe amnloLlc cavlLy. LxploraLlon of Lhe amnloLlc cavlLy and Lhe feLus
was done afLer lrrlgaLlon of 3 Lo 10mL normal sallne soluLlon aL body LemperaLure. Alms
of Lhe feLoscoplc procedure were Lo ldenLlfy aL leasL Lhe feLal head (ears, mouLh or
nose, llg 2), Lwo exLremlLles (llg 3), Lhe Lall, and umblllcal cord and/or Lhe placenLa.
Maxlmum feLoscopy duraLlon was seL aL Len mlnuLes. AfLer Lhe procedure Lhe uLerus
was closed and reposlLloned and Lhe abdomen closed ln layers. AL 30 days of CA Lhe
anlmals were euLhanlzed Lo undergo evaluaLlon of feLal survlval and welghL.
41

8LSuL1S: 1he needle could always be correcLly poslLloned lnLo Lhe amnloLlc cavlLy, uslng
Lhe mlcroscopy gulded open access Lechnlque. 8y lnfuslon of max. 10mL of normal
sallne, sufflclenL work space could be creaLed. leLoscoplc lnspecLlon of feLal head (ears,
mouLh or nose), exLremlLles, back, Lall, and umblllcal cord could be done, as well as
membrane and placenLa was compleLe ln all lnsLances. 8esoluLlon was [udged Lo be
adequaLe: Lhe vlew was sharp, and small feLal cuLaneous or membraneous blood vessels
could be easlly vlsuallzed. Cccaslonally feLal movemenLs and degluLlLlon were seen. no
feLal or lnLra-amnloLlc haemorrhage occurred. no posLoperaLlve maLernal deaLhs,
lnfecLlons or aborLlons were noLed ln Lhls serles. leLal survlval raLe was 100 ln Lhe
sLudygroup (feLoscopy group) and 83.7. ln negaLlve conLrols (p>0.03). Mean welghL
of llve born feLuses was 36.3!3.27gr. ln Lhe sLudygroup and 33.2!1.48gr. ln negaLlve
conLrols (p=0.2983). no lnLra-amnloLlc nor lnLra-abdomlnal adheslons were found aL Lhe
second look operaLlon.
ulSCuSSlCn: 1here has been renewed lnLeresL for endoscoplc access Lowards Lhe feLus.
We have proposed a sheep model for feLoscoplc surgery, whlch was used for urlnary
LracL obsLrucLlon and foreguL endoscopy. LxperlmenLal work has also been done on
Lracheal obsLrucLlon, Lhe lnserLlon of a bladder sLenL or mlcrosuLurlng 3. Powever, Lhe
sheep model ls expenslve, and gesLaLlon ls lengLhy. lor some purposes, a smaller anlmal
model may be useful. 1hls new, lnexpenslve, readlly avallable anlmal model, whlch does
noL requlre any speclal faclllLles for lodglng or anaesLhesla, could conLrlbuLe Lo Lhe
developmenL of feLoscopy. lL may even be used for surglcal manlpulaLlons on Lhe feLus,
placenLa or membranes. Moreover Lhe rabblL has a blcornaLe and qulescenL uLerus wlLh
many amnloLlc sacs and feLuses. 1hese feLuses are large enough Lo be surglcal
manlpulaLed as Longaker eL al. demonsLraLed 6. We suggesL Lo use Lhls model for
Lralnlng ln feLoscopy, and perhaps for surglcal research, such as feLal wound, bone and
nerve heallng, feLal gene Lherapy or more surglcal condlLlons as clefL llp and splnal cord
leslon ln feLal plasLlc surgery. We earller used Lhe rabblL model Lo sLudy Lhe heallng
process of Lhe feLal membranes followlng hysLeroLomy 4 and have now embarked on
Lhe sLudy of feLoscopy-lnduced membrane Lrauma. ln concluslon, ln Lhe feLal rabblL
model, excellenL feLoscoplc vlsuallzaLlon could be obLalned wlLhouL obvlous feLal slde
effecLs.

42

3D computer-tomograph|c cepha|ometr|c eva|uat|on of |ntrauter|ne repa|red c|eft-||ke
defects
apadopoulos nA,
1
apadopoulos MA,
2
!annowlLz 1,
3
8oeLLcher ,
3
kloeppel M,
3
Penke
!,
3
Mueller u,
1
kovacs L,
1
Zellhofer P-l,
4
8lemer L
1
1
uepL. of lasLlc and 8econsLrucLlve Surgery, 1echnlcal unlverslLy of Munlch, Cermany,
2
uepL. of CrLhodonLlcs, School of uenLlsLry, ArlsLoLle unlverslLy of 1hessalonlkl, Creece,
3
lnsLlLuLe of LxperlmenLal Cncology and 1herapy 8esearch, 1echnlcal unlverslLy of
Munlch, Cermany, 4CenLer of Advanced SLudles ln Cranlo-Maxlllofaclal Surgery,
1echnlcal unlverslLy of Munlch, Cermany & uepL. of Cral & Maxlllofaclal Surgery,
unlverslLy of 8asel, SwlLzerland.

ln18CuuC1lCn: ClefL llp wlLh or wlLhouL clefL palaLe ls one of Lhe mosL common sLrucLural blrLh
defecLs, whlch managemenL requlres a mulLldlsclpllnary approach, lncludlng mulLlple surgerles
over Lhe flrsL 18 years of llfe. All surglcal lnLervenLlons have an lmpacL on Lhe cranlofaclal growLh
of Lhe young paLlenL and Lhelr consequences dlffer accordlng Lo Lhe exLenL of Lhe clefL and Lhe
Lechnlques used for lLs correcLlon. LxperlmenLal lnLrauLerlne LreaLmenL of clefL llp and palaLe
has capLured Lhe lnLeresL of cranlofaclal surgeons, afLer lL was observed LhaL Lhrough an
lnLervenLlon aL Lhe mldgesLaLlonal age, sofL Llssue wound heallng could occur wlLhouL scars and
bone heallng could Lale place wlLhouL or wlLh mlnlmal callus formaLlon. 1hls way, prlmary or
secondary maxlllary growLh resLrlcLlons could be mlnlmlzed, resulLlng probably ln a reducLlon or
mlnlmal need of secondary surglcal lnLervenLlons or addlLlonal LreaLmenLs afLer blrLh, lncludlng
orLhodonLlcs. ConsequenLly, durlng Lhe lasL Lwo decades dlfferenL anlmal models have been
proposed Lo evaluaLe lnLrauLerlne LreaLmenL of clefL llke defecLs. unforLunaLely, Lhese sLudles
were deallng only wlLh Lhe malformed upper llp sofL Llssues and wlLh wound heallng. 1he bony
alveolar defecL, whlch has been slmulLaneously creaLed remalned unLreaLed and was noL
consldered, alLhough some buL noL compleLe sponLaneous closure could occur. 1hls slLuaLlon,
when ln addlLlon some form of callus formaLlon Lake place, can lead Lo resLrlcLlon of growLh of
Lhe correspondlng parL of Lhe maxllla, and Lherefore Lo maxlllary deflclency and asymmeLrles.
AlM: 1he alm of Lhls sLudy was Lo evaluaLe lnLrauLerlne reconsLrucLlon of clefL-llke defecLs ln Lhe
mld-gesLaLlonal sheep model by means of Lhree-dlmenslonal compuLer-Lomographlc
cephalomeLrlc analysls.
MA1L8lAL Anu ML1PCuS: 1he creaLlon of clefL llp and alveolar llke defecLs, grafL harvesLlng,
repalr of clefL llp and alveolar llke defecLs, as well as closure of Lhe uLerus were per- formed
beLween Lhe 73Lh and 93Lh gesLaLlon day (Lerm: 143-130 days). 1welve feLuses were used for
Lhe surglcal creaLlon of a unllaLeral CLA llke defecL and lLs subsequenL ln uLero repalr, whlch
were asslgned ln 4 sLudy groups. AfLer laparoLomy and hysLeroLomy and keeplng Lhe head of fe-
Luses ouL of Lhe uLerus, a unllaLeral clefL llp wlLh an alveolar defecL of clrca 2 mm wldLh was
creaLed (llg. 1). lor Lhe closure of Lhe alveolar defecL, feLal lllac cresL bone grafL was used by Lhe
feLuses of group l (n=3), whlch was flxed Lo Lhe maxllla wlLh flbrln sealanL (1lssucol , 8axLer
Cermany - lmmuno CmbP, Peldelberg, Cermany). 8y Lhe feLuses of group ll (n=2), a feLal ulnar
grafL was used for Lhe alveolar defecL closure, whlch was also flxed Lo Lhe maxllla wlLh flbrln
43

sealanL (1lssucol ). A feLal ulnar grafL was used also by Lhe feLuses of group lll (n=3), whlch was
flxaLed Lo Lhe maxllla as ln group ll, Lhus comblned wlLh a Core-1ex membrane (186? Core-1ex
8egeneraLlve MaLerlal - membrane - 1lLanlum-8elnforced , W.L. Core & AssoclaLes CmbP,
uLz- brunn, Cermany). 8y Lhe feLuses of group lv (n=4), Lhe bone regeneraLlng bloresorbable
maLerlal collagen-lyophlllsaLe (Coloss Collagen-LyophlllsaLe ) covered also by Lhe Core-1ex
membrane, was used for Lhe closure of Lhe surglcally creaLed alveolar defecL. 1he unLreaLed
conLrol group was conslsLed of 4 feLuses. llnally, muscle adapLaLlon of Lhe upper llp and skln
closure were per- formed. AfLer Lhe end of manlpulaLlon, Lhe feLal heads were placed back lnLo
Lhe uLerus (llg. 2). AL Lhe 140-143 gesLaLlon day Lhe experlmenLal as well as Lhe conLrol anlmals
underwenL a second look operaLlon and euLhanasla. AfLer euLhanasla, a C1-scannlng of each
skull of Lhe operaLed feLuses was performed and a Lhree-dlmenslonal reconsLrucLlon by means
of Lhe vCx- LLMAn sofLware was followed. llnally, a meLhod of Lhree-dlmenslonal
cephalomeLrlc analysls was developed for Lhe purposes of Lhe sLudy (llg. 3 & 4). 1welve
varlables were used. 1he sLaLlsLlcal analysls was performed by means of Lhe SSS sofLware, and
lncluded lnLra-group comparlsons beLween Lhe operaLed and non-operaLed skull-sldes, and
lnLer-group comparlsons of Lhe lefL/rlghL skull slde asymmeLry.
8LSuL1S: 1he lnLra-group comparlsons beLween operaLed and non-operaLed skull- sldes
revealed no sLaLlsLlcal slgnlflcanL dlfferences for Lhe 12 varlables used, ln each group separaLely
(llg. 3) uue Lo Lhe small number of feLuses per group and ln order Lo lncrease Lhe value of Lhe
sLaLlsLlcal evaluaLlon, we grouped all Lhe operaLed feLuses LogeLher, and we performed Lhe
same comparlsons beLween Lhe Lwo sldes. ln Lhls case, Lhree of Lhe Lwelve cephalomeLrlc
varlables presenLed slgnlflcanL dlfferences (llg. 6). 1aklng lnLo accounL LhaL bone grafL heallng
could be dlagnosed only ln half of all Lhe operaLed feLuses, we LreaLed Lhem as a separaLe group
and com- pared also Lhelr Lwo skull sldes. ln Lhls case, none of Lhe varlables revealed slgnlflcanL
dlfferences (llg. 7). 1he lnLer-group comparlsons of Lhe lefL/rlghL skull slde asymmeLry be-
Lween all Lhe operaLed feLuses and Lhe conLrol group revealed LhaL half of Lhe cephalomeLrlc
varlables presenLed slgnlflcanL dlfferences (llg. 8). lrom Lhe oLher hand, when we compared Lhe
lefL/rlghL asymmeLry beLween Lhe feLuses LhaL bone grafL heallng Look place and Lhe conLrol
group, only one varlable dlffered slgnlflcanLly (llg. 9). Some represenLaLlve 3u-C1 skull
reconsLrucLlons from anlmals from Lhe dlfferenL groups are shown on llgures 10-14.
CCnCLuSlCnS: Accordlng Lo Lhls sLudy, each of Lhe four lnLrauLerlne LreaLmenL meLhods seems
noL Lo affecL slgnlflcanLly Lhe posL-surglcal maxlllary growLh. no sLaLlsLlcal slgnlflcanL dlfferences
could be demonsLraLed beLween Lhe anlmals wlLh bone heallng and Lhese wlLhouL bone heallng,
alLhough, a Lendency of superlor normal growLh of Lhe premaxllla could be observed, when
bone heallng occurs.
44

MSC-I|||ed Surg|ca| Sutures to romote Wound nea||ng

8eckhenrlch Ak
1
, klrsch 8M
1
, Wahl LA
1
, Schenck 1L
1
, 8ezaelan l
1
, Parder ?
1
, loehr
3
,
Machens PC
1
and Lgana !1
1,2

1
ueparLmenL of lasLlc and Pand Surgery, kllnlkumrechLs der lsar, 1echnlsche
unlverslLaL Munchen, Munlch, Cermany.
2
lCnuA CenLer forCenome 8egulaLlon,
laculLad de Clenclas, unlversldad de Chlle, SanLlago, Chlle.
3
Chalr for CrLhopaedlcs and
SporL CrLhopaedlcs, kllnlkumrechLs der lsar, 1echnlsche unlverslLaL Munchen, Munlch,
Cermany.

uelayed wound heallng and scar formaLlon are among Lhe mosL frequenL compllcaLlons
afLer surglcal lnLervenLlons. AlLhough blodegradable surglcal suLures presenL an
excellenL drug dellvery opporLunlLy, Lhelr prlmary funcLlon ls flxaLlon. Mesenchymal
sLromal cells (MSCs) acL as Lrophlc medlaLors and are successful ln acLlvaLlng
blomaLerlals. Pere blodegradable suLures were fllled wlLh adlpose-derlved MSCs Lo
provlde a pro-regeneraLlve envlronmenL aL Lhe ln[ured slLe. 8esulLs showed LhaL afLer
fllllng, MSCs aLLach Lo Lhe suLure maLerlal, dlsLrlbuLe equally LhroughouL Lhe fllamenLs
and remaln vlable ln Lhe suLure. Among a broad panel of cyLoklnes, fllled suLures
consLanLly release vascular endoLhellal growLh facLor. Such condlLloned medla was
evaluaLed ln an ln vlLro wound heallng assay and showed a slgnlflcanL decrease ln Lhe
open wound area compared Lo conLrols. AfLer suLurlng ln an ex vlvo wound model cells
remalned ln Lhe suLure and malnLalned Lhelr meLabollc acLlvlLy. lurLhermore, cell-fllled
suLures provlde Lhe ablllLy Lo be cryopreserved. 1hls sLudy presenLs an lnnovaLlve
approach Lo equlp surglcal suLures wlLh pro-regeneraLlve feaLures and allows Lhe
LreaLmenL and flxaLlon of surglcal wounds ln one sLep. 1herefore MSC-fllled suLures are
a promlslng Lool Lo promoLe wound heallng afLer surgery and could avold poor scar
formaLlon.



43

Lp|genet|c contro| of the Cster|xgene promoter act|v|ty dur|ng mesenchyma|ce||
d|fferent|at|on
Pugo Sepulveda, Candy 8o[as and MarLln MonLeclno.
CenLer for 8lomedlcal 8esearch and lCnuA CenLer for Cenome 8egulaLlon, laculLy of
8lologlcal Sclences and laculLy of Medlclne, Andres 8ello unlverslLy, Chlle.

CsLerlx (Csx) ls a key regulaLor of osLeoblasL dlfferenLlaLlon as lL conLrols LranscrlpLlon of
a number of bone-phenoLyplcgenes. Pere, we lnvesLlgaLe eplgeneLlc mechanlsms LhaL
govern Csx gene LranscrlpLlon durlng Lhe engagemenL ofplurlpoLenL mesenchymal cells
Lo Lhe osLeoblasL llneage. LplgeneLlc posLLranslaLlonal modlflcaLlons and blndlng
ofregulaLory proLelns Lo chromaLln were deLermlned by chromaLln
lmmunopreclplLaLlon. unA meLhylaLlon was deLermlned bycleavage wlLh meLhylaLlon-
senslLlve resLrlcLlon endonucleases and lmmunopreclplLaLlon of meLhylaLed unA.
Changes lnCsx m8nA were assessed by 81-qC8. 1he effecL of Lhe hlsLone-deaceLylase
(PuAC) lnhlblLor 1SA was verlfled byWesLern bloL. We flnd LhaL Lhe Csx gene promoLer
lnosLeoblasLlc cells exhlblLs hlsLone modlflcaLlons normally assoclaLedwlLh
LranscrlpLlonal acLlvlLy (P3Ac/P3k4me3) LogeLher wlLh mlnlmal unA meLhylaLlon. ln
conLrasL, non-bone cells showhlsLone modlflcaLlons assoclaLed wlLh LranscrlpLlonal
represslon (P3k9me3/P3k27me3) and elevaLed unA meLhylaLlon.lnLeresLlngly,
uncommlLLed plurlpoLenL mesenchymal cells show a comblnaLlon of boLh Lypes of
hlsLone modlflcaLlons(P3k4me1/P3k27me3), accompanled by reduced, buL sLlll
slgnlflcanL, unA meLhylaLlon. CommlLmenL of Lhese cells LoelLher bone or muscle
llneages ls accompanled by changes ln chromaLln modlflers lncludlng PuACs, hlsLone-
meLhylLransferases(PM1s) and unA-meLhylLransferases (unM1s) aL Lhe Csx promoLer ln
LlghL correlaLlon wlLh LheLranscrlpLlonal sLaLus. unM11, unM13a, PuAC2, and PuAC4
are enrlcheddurlng Csx sllenclng ln muscle cells, whereasLhese proLelns are released
from Lhe Csx promoLerdurlng osLeoblasL dlfferenLlaLlon, LogeLher wlLh demeLhylaLlon of
unA.Also, lnhlblLlon of PuAC and unM1 acLlvlLles ln uncommlLLed mesenchymal
precursors lnduces Csx m8nA expresslon.1hus, PlsLone and unA modlflcaLlons
represenL key regulaLory mechanlsms durlng conLrol of Csx gene expresslon
lnmesenchymal-derlved cell llneages, Lhereby conLrolllng Lhe osLeoblasLlc cell faLe
cholce.
SupporLed by CCnlC?1 uocLoral lellowshlps rogram, lCnuA 13090007, lCnuLC?1
1093073, unA8 ul-77-12/l

46

In V|tro Lva|uat|on of Scaffo|ds for the De||very of Mesenchyma| Stroma| Ce||s
1hllo Schenck
1
, LllzabeLh Wahl
1
, lernando llerro
2
, 1homas eavy
3
, Pans-CunLher
Machens
1
, ursula Popfner
1
, Manuela klrsch
1
, !ullan uye
4
, larld 8ezaelan
1
and!. 1oms
Lgana
1,3

1
ueparLmenL of lasLlc Surgery and Pand Surgery, kllnlkum rechLs der lsar, 1echnlsche
unlverslLaL Munchen, Munlch, Cermany.
2
lnslLuLe for 8egeneraLlve Cures, uC uavls, CA,
uSA.
4
8Al1 lnsLlLuLe, London, uk,
3
londap CenLer for Cenome 8egulaLlon, laculLad de
Clenclas, unlversldad de Chlle, SanLlago, Chlle.

Mesenchymal sLromal cells (MSCs) have shown Lo lmprove Llssue regeneraLlon ln
several pre- and cllnlcal Lrlals. 1hese cells have been used ln comblnaLlon wlLh Lhree-
dlmenslonal scaffolds as a promlslng approach ln Lhe fleld of regeneraLlve medlclne and
Lhe sLudy of such lnLeracLlons has becomlng a key lssue ln Lhe fleld of blomaLerlals. ln
Lhls work, we compare Lhe behavlour of MSCs afLer seedlng onLo four dlfferenL
blomaLerlals, lncludlng some LhaL are commonly used ln cllnlcal seLLlngs. MSCs derlved
from human faL Llssue were lsolaLed, characLerlzed, and seeded onLo scaffolds
consLrucLed malnly of collagen, flbrln, chlLosan, or decellularlzed dermls. 8esulLs
showed LhaL Lhe scaffolds sLrongly lnfluences key parameLers, such as, seedlng
efflclency, cellular dlsLrlbuLlon, aLLachmenL, survlval, meLabollc acLlvlLy, and paracrlne
release. AlLhough furLher ln vlvo sLudles have Lo be performed, Lhls work provldes
lmporLanL lnformaLlon for cllnlcal LranslaLlon and represenLs a sLep for opLlmlzlng Lhe
use of MSC-seeded scaffolds ln Llssue regeneraLlon.

47

1he deve|opment of a t|ssue eng|neered |ntegrated |mp|ant system for the treatment
of art|cu|ar cart||age defects
1, 4
SoLo C,
2
8rown u,
4
Cerlanl 8,
3
Albornoz l,
3
Snchez L,
3
?oung M and
4
WelnsLeln-
Cppenhelmer C.
1
CenLro de 8egulacln del Cenoma, laculLad de Clenclas, unlversldad de
Chlle,
2
ueparLamenLo de 8lologla, LaboraLorlo de 8lologla de la 8eproduccln y el
uesarrollo,laculLad de Clenclas, unlversldad de valparalso,
3
CenLro de 8loLecnologla
uanlel Alakalay, unlversldad 1ecnlca lederlco SanLa Marla,
4
ueparLamenLo de
8loqulmlca, laculLad de larmacla, unlversldad de valparalso.

CarLllage defecLs represenL a blg cllnlcal lssue and mlghL be caused by LraumaLlc ln[ury
or some oLher facLors LhaL compromlse Lhe hyallne arLlcular carLllage. 1o daLe no
LreaLmenLs are avallable Lo fully recover carLllage regeneraLlon. Powever, wlLhln Lhe lasL
years, Llssue englneerlng has offered a new and promlslng LherapeuLlc opLlon for Lhe
LreaLmenL of hyallne arLlcular carLllage defecLs.
ln Lhls sLudy we developed an lnLegraLed lmplanL SysLem for Lhe LreaLmenL of arLlcular
carLllage defecLs. Pere rabblL arLlcular chondrocyLes were lncorporaLed and lmmoblllzed
ln a 3u scaffold composed of gelaLln, chlLosan, hyaluronlc acld, and a flbrln. AfLer
seedlng, cell vlablllLy and prollferaLlon of Lhe chondrocyLes was examlned aL dlfferenL
Llmes. uslng blochemlcal and lmmunohysLochemlcal meLhods, resulLs showed a
slgnlflcanL lncrease on cell vlablllLy (p < 0,03) and prollferaLlon (p < 0,03) over Llme.
lnLeresLlngly, boLh parameLers were slgnlflcanLly lncreased ln Lhe lmplanL when
compared Lo chondrocyLes culLured ln convenLlonal monolayer culLure.
Cur resulLs show LhaL such lnLegraLed lmplanL SysLem ls a promlslng carrler Lo dellver
arLlcular chondrocyLes for Lhe LreaLmenL of carLllage defecLs.

48

Iso|at|on of Mesenchyma| Stem Ce||s (MSC) from Lqu|ne Amn|on: A otent|a| Source
to 1reat norse Ske|eta| Musc|e In[ur|es
n. SLephens
1
, Ml CrLuzar
1
, S. San MarLln
2.3
, M. rraga
2.3
, A. Moller
3
and !. vlllena
2.3
1
Porse & Well, Lqulne Allogenlc 8egeneraLlve Medlclne,
2
8lomedlcal 8esearch CenLre,
laculLy of Medlclne, unlversldad de valparalso, Chlle,
3
lnblocrloLec S.A
Lqulne amnlon represenLs an aLLracLlve source Lo obLaln MSC. Among oLhers, amnlon ls
a blg and young Llssue LhaL do noL requlre an lnvaslve procedure Lo be obLalned and lL
uses do noL ellclL eLhlcal debaLe. 1he alm of Lhls work was Lo esLabllsh a rellable proLocol
Lo lsolaLe and characLerlze MSC obLalned from equlne amnlon.
1en amnlon samples were collecLed from Lwo mares and Lhelr Llssue orlgln was
conflrmed by hlsLologlcal analyses. AfLerwards samples were sub[ecLed Lo enzymaLlc
dlgesLlon (1rlpsln-Lu1A soluLlon) and culLured lnLplllfe
1M
and lL was replaced every
seconddays.8esulLs showed hlgh cell denslLles wlLh vlablllLy of aL leasL 80. AfLerwards
cells were characLerlzed and resulLs show LhaL Lhe lsolaLed cells share common feaLures
wlLh MSC lsolaLed from oLher sources.
1he maln drawback of Lhls proLocol ls LhaL 80 of Lhe samples were conLamlnaLed wlLh
bacLerlal and fungal, due Lo Lhe envlronmenL where Lhe procedure of amnlon sampllng
was performed. 1hus new proLocols are currenLly under evaluaLlon.
luLure approaches conslder Lhe use of such cells for Lhe LreaLmenL of horse skeleLal
muscle ln[urles ln allogenelc procedures.

keywords: Amnlon, equlne, mesenchymal sLem cells.

49

Immune gene express|on |n response to sp|na| cord |n[ury |n "#$%&'()*#+,(
vlcLor 1apla, Maurlclo Moreno, uasfne Lee and !uan Larraln
CenLer for Aglng and 8egeneraLlon and Mlllenlum nucleus ln 8egeneraLlve 8lology

ln conLrasL Lo mammals, \eooposloevls Ladpoles are able Lo regeneraLe Lhe splnal cord
afLer ln[ury, buL Lhls regeneraLlve capaclLy ls losL durlng meLamorphosls. Cur alm ls Lo
deLermlne lf Lhere ls a dlfferenLlal expresslon of lmmune genes ln response Lo splnal
cord ln[ury (SCl) comparlng regeneraLlve and non-regeneraLlve sLages of \. loevls.
SCl was done by LransecLlon on pre- and posL-meLamorphlc sLages. AfLer 1, 2 and 6 days
posL-LransecLlon (dpL) LoLal 8nA was exLracLed from Lhe splnal cord. A hlgh-LhroughpuL
8nA sequenclng analysls (8nA-Seq) was performed and m8nA levels were quanLlfled by
81-qC8. lurLhermore, gene expresslon was characLerlzed afLer SCl by lo slto
hybrldlzaLlon (lSP) and proLeln level by lndlrecL lmmunofluorescence.
uaLa from 8nA-Seq analysls lndlcaLes LhaL ln response Lo SCl Lhere ls a dlfferenLlal
lmmune response. ln regeneraLlve sLages genes assoclaLed Lo lmmunlLy are regulaLed Lo
a basal level afLer 6 dpL, buL ln non-regeneraLlve sLage genes are sLlll up-regulaLed. We
valldaLed Lhls analysls for several genes uslng dlfferenL experlmenLal approaches: some
genes were valldaLed by 81-qC8, ln addlLlon Lhe dlfferenLlal levels of MPCll complex
was valldaLed by lmmunofluorescence and for lasL, Lhe dlfferenLlal expresslon of SCCS3,
LargeL for Lhe !ak-SLaL paLhway, was characLerlzed by lSP.
Cur resulLs lead us Lo correlaLe Lhe dlfferenLlal lmmune response wlLh Lhe loss of Lhe
regeneraLlve capaclLy of \. loevls. As long-Lerm ob[ecLlve ls expecLed Lo undersLand how
lnflammaLlon, whlch ls non-resolvlng and has deLrlmenLal effecLs on SCl ln mammals,
affecLs Lhls regeneraLlve process ln \eoopos.

Grants londecyL 11100348, Mln8L8 07/011, CA8L

30

Dynam|cs of degenerat|on and regenerat|on |n deve|op|ng zebraf|sh per|phera| axons
revea|s a requ|rement for extr|ns|c ce|| types.

vlllegas 8, MarLln SM, C'uonnell kC, Carrlllo SA, SagasLl A and Allende ML.
lCnuA CenLer for Cenome 8egulaLlon, laculLad de Clenclas, unlversldad de Chlle.

undersLandlng Lhe cellular mechanlsms regulaLlng axon degeneraLlon and regeneraLlon
ls cruclal for developlng LreaLmenLs for nerve ln[ury and neurodegeneraLlve dlsease. ln
neurons, axon degeneraLlon ls dlsLlncL from cell body deaLh and ofLen precedes or ls
assoclaLed wlLh Lhe onseL of dlsease sympLoms. ln Lhe perlpheral nervous sysLem of
boLh verLebraLes and lnverLebraLes, afLer degeneraLlon of deLached fragmenLs, axons
can ofLen regeneraLe Lo resLore funcLlon. Many sLudles of axonal degeneraLlon and
regeneraLlon have used ln vlLro approaches, buL Lhe lnfluence of exLrlnslc cell Lypes on
Lhese processes can only be fully addressed ln llve anlmals. 8ecause of lLs slmpllclLy and
superflclal locaLlon, Lhe larval zebraflsh posLerlor laLeral llne (pLL) nerve ls an ldeal
model sysLem for llve sLudles of axon degeneraLlon and regeneraLlon.


31

numan Wharton's Ie||y Mesenchyma| Stem Ce||s (WI-MSC) promotes ang|ogenes|s
both ,$!+,-.%!and ,$!+,+%!and have potent|a| therapeut|c propert|es.

Zavala C
1
, Ldwards S
1
, rleLo C
1
, CrLlz C
1
, MarLlnez S
1
, Lgana !1
2
, 8ono M8
3
and alma v
1
1
LaboraLory of SLem Cells & uevelopmenL, laculLy of Sclences, unlversldad de Chlle,
Chlle,
2
LaboraLory of 1lssueLnglneerlng&8egeneraLlon, 1echnlcalunlverslLy of Munlch,
Cermany,
3
LaboraLory of lmmunology, laculLy of Sclences, unlversldad de Chlle, Chlle.

Introduct|on. W!-MSC are a mesenchymal populaLlon of cells lsolaLed from Lhe
gelaLlnous subsLance LhaL surround blood vessels ln Lhe umblllcal cord. As MSC, Lhese
cells possess prollferaLlon and dlfferenLlaLlon capablllLles buL also, W!-MSC secreLe a
dlverse and rlch seL of cyLoklnes and growLh facLors. Some of Lhese growLh facLors have
reporLed pro-anglogenlc properLles, Lherefore we wanLed Lo evaluaLe lf W!-MSC have
Lhe poLenLlal Lo promoLe new blood vessels formaLlon Lhrough secreLlon of pro-
anglogenlc molecules. W!-MSC and Lhelr condlLloned medlum (CM) were assayed lo
vltto Lhrough Lubule formaLlon assay wlLh Puman umblllcal veln LndoLhellal Cells
(PuvLC) and lo vlvo ln a mouse model for scaffold-based dermal anglogenesls.
Mater|a| and methods. W!-MSC and PuvLC were lsolaLed from normal umblllcal cords.
W!-MSC were sLlmulaLed Lo dlfferenLlaLe Lo adlpose and bone Llssue uslng speclflc
llneage-lnducLlve medlums, afLer 2 weeks cells were dyed wlLh 8ed Cll C and Allzarln
8ed Lo demonsLraLe dlfferenLlaLlon Lo adlpose and bone Llssue, respecLlvely. ln addlLlon
W!-MSC were characLerlzed by Lhe expresslon of Lhe mesenchymal markers Cu73,
Cu103, Cu90, Cu44 and Cu29 and negaLlve expresslon of Lhe hemaLopoleLlc markers
Cu43, Cu14 and PLA-u. CM was generaLed wlLhouL serum for 48 hours and used Lo
evaluaLe Lhe conLenL of anglogenlc molecules byC8, WesLern bloL and roLeome
profller array. PuvLC were sLlmulaLed wlLh CM beLween 4-6 hours and Lubule formaLlon
assay was evaluaLed by counLlng Lubules number and ramlflcaLlon polnLs. ln order
LoevaluaLe anglogenesls ln mlce, W!-MSC were seeded on scaffolds (12 mm, lnLegra
MaLrlx) for 24 hours, followed by suLuraLlon Lo Lhe wounded back of 8AL8C mlce. AfLer
4-10 days, mlce were sacrlflced and anglogenesls was evaluaLed by lmage analysls,
quanLlfylng Lhe vascular neLwork ln Lhe scaffold. ln addlLlon, lnflammaLlon was analyzed
32

Lhrough esLlmaLlon of Lhe lnflammaLory cyLoklne conLenL of blood plasma by C8A
(cyLoklne bead array).
kesu|ts. W!-MSC are ldenLlfled as amesenchymal populaLlon of cells because Lhey have
Lhe poLenLlal Lo dlfferenLlaLe Lo adlpose and bone Llssue afLer 2 weeks of sLlmulaLlon,
express classlc markers of MSC and are negaLlve for hemaLopoleLlc markers. W!-MSC
secreLe a dlverse seL of cyLoklnes and growLh facLors wlLh anglogenlcpoLenLlalsuch as
vLCl, lCl-2, uCl, Anglogenln among oLhers. PuvLC LreaLed wlLh CM ln Lubule
formaLlon assayformed Lubules afLer 4 hours, ln comparlson wlLh Lhelr negaLlve conLrol
(uMLM). AddlLlonally, ln mlce model W!-MSC can lnduce new vessel formaLlon lnLo Lo
Lhe scaffold wlLh respecL Lo empLy scaffolds afLer 4-10 days of LreaLmenL.
Conc|us|ons. W!-MSC are aLLracLlve for regeneraLlve Lherapy because Lhey secreLe
anglogenlc facLors and can promoLe vessel formaLlon by acLlng boLh on endoLhellal cells
(PuvLC) and blood vessels (mouse anglogenesls assay), whlch shows LhaL boLh lo vltto
ooJ lo vlvo, W!-MSCs can secreLe facLors wlLh LherapeuLlc poLenLlal.





33


Contact Informat|on
I|rst Ch||ean - German Workshop 2013

8odrlgo Agullar ragullarmaurelra[hoLmall.com unlversldad Andres 8ello, Chlle
Mlguel Allende allende[uchlle.cl unlversldad de Chlle, Chlle
Mar[orle lvarez mar[orl.alvarez[gmall.com unlversldad de Chlle, Chlle
!ose lgnaclo Arlas [oarlas[uchlle.cl unlversldad de Chlle, Chlle
Marcelo Campos faclen03[uchlle.cl unlversldad de Chlle, Chlle
Myra Chvez myra.chavez.rosas[Lum.de 1echnlsche unlverslLaL Munchen, Cermany
!ullan uye dye[[rafL.ac.uk 8Al1 lnsLlLuLe, MounL vernon PosplLal,
Mlddlesex, London, uk.
Cabrlela Ldwards gabrlela.edwards[gmall.com . unlversldad CaLllca, Chlle
1oms Lgana Lomasega[gmall.com 1echnlsche unlverslLaL Munchen, Cermany
AlberLo llerro aflerro[cllnlcalascondes.cl Cllnlca Las Condes, Chlle
lernando llerro feflerro[gmall.com unlverslLy of Callfornla aL uavls, uSA
?ves Parder harder[lrz.Lu-muenchen.de 1echnlsche unlverslLaL Munchen, Cermany
Claudlo PeLz claudlo.heLz[gmall.com unlversldad de Chlle, Chlle
Cudrun kausel gkausel[gmall.com unlversldad AusLral, Chlle
Maroun khoury Mkhoury[uandes.cl unlversldad de Los Andes, Chlle
!uan Larraln [larraln[blo.puc.cl onLlflcla unlversldad CaLllca, Chlle
Serglo Lavandero slavander[uchlle.cl unlversldad de Chlle, Chlle
Pans-CunLher Machens machens[lrz.Lu-muenchen.de 1echnlsche unlverslLaL Munchen, Cermany
naLalla Mackenzle naLalla.mackenzle[gmall.com unlversldad Andres 8ello, Chlle
MarLln MonLeclno mmonLeclno[unab.cl unlversldad Andres 8ello, Chlle
Andreas nussler andreas.nuessles[googlemall.com Lberhard- karls unlverslLaL 1ublngen,
Cermany.
vernlca alma vpalma[uchlle.cl unlversldad de Chlle, Chlle
norberL allua npallua[ukaachen.de unlverslLy PosLplLal 8W1P, Aachen,
Cermany
34

nlkolaos apadopulos n.papadopulos[lrz.Lu-muenchen.de 1echnlsche unlverslLaL Munchen, Cermany
Ann 8eckhenrlch ann.reckhenrlch[lrz.Lu-
muenchen.de

1echnlsche unlverslLaL Munchen, Cermany
Pugo Sepulveda hugosepulvedalnosLroza[gmall.com unlversldad de Chlle, Chlle
1hllo Schenck 1hllo.Schenck[lrz.Lu-muenchen.de 1echnlsche unlverslLaL Munchen, Cermany
Claudlo SoLo claudlosoLo.valenzuela[gmall.com unlversldad de Chlle, Chlle
SebasLln San MarLln sebasLlan.sanmarLln[uv.cl unlversldad de valparalso, Chlle
naLalla SLephens naLallasLephens.s[gmall.com Porse & Well, Chlle
vlcLor 1apla vlcLor.Lapla.o[gmall.com . unlversldad CaLllca de valparalso, Chlle
8osarlo vlllegas rosarlovs[gmall.com . unlversldad CaLllca, Chlle
Cabrlela Zavala gzavalavlvar[gmall.com unlversldad de Chlle, Chlle






33

Ad|t|ona| Informat|on



lf you would need any klnd of help please flnd here Lhe moblle numbers of:
Mlguel Allende: 9 - 6 - 691 03 29
1omas Lgana: 9 - 6 - 692 07 94




lf you are looklng for a hoLel Lo sLay we recommend some:
hLLp://renLasulLe.cl/
hLLp://www.plazaelbosque.cl/parkandsulLes/foLoLour.hLml
hLLp://www.nh-hoLels.com/nh/en/hoLels/chlle/sanLlago-de-
chlle/nh-cludad-de-sanLlago.hLml
hLLp://elbosque.aLLon.com/defaulL-es.hLml

36

Sant|ago subway
1he workshop wlll be aL Lhe meLro Un|vers|dad Cat||ca" over Lhe red llne.




37

Workshop rooms
Cnce you arrlve aL Lhe . unlversldad CaLllca de Chlle (above) Lake Lhe flrsL enLrance
(Alameda 324) and you wlll be aL Lhe lotlo Je lo vltqeo (below). 1he workshop wlll be
on Lhe 2nd floor (5olo Je nooot).



38






















www.genomacrg.c|
I|rst Ch||ean - German Workshop
Deve|opmenta| 8|o|ogy and kegenerat|ve Med|c|ne: from t|ssue generat|on to re - generat|on

londap CenLer for Cenome 8egulaLlon
Av. 8lanco Lncalada 2083, 3
rd
floor
SanLlago de CPlLL