Part I: HK Stem Cell Research - - - Facts and Fallacies.

By IAAM President Dr. Hans J. Kugler, PhD

DrKugler@roadrunner.com

Part I: Stem Cell Facts and Fallacies. Part II: Frequently asked questions. Part III: Pictures of our stem cell lab. Part IV: Parthenogenesis, Virgin-Birth.

Very recently on June 6, 2006, Harvard University announced that they are initiating stem cell research to make their own lines of stem-cells. A few days later on June 11, 2006 in the Los Angeles Times, we learned that "Stem Cell Labs Take Private Path." Two privately funded New York laboratories are conducting their own stem cell research. Both of the above avenues of stem cell research are directed to nding better treatments for a wide spectrum of diseases, including but not limited to Lou Gehrig's, Parkinsons, diabetes, cancer and anti-aging.

IAAM is following a similar path. Our laboratory is fully equipped and research has already begun (see pictures on our web-site). At this time, we invite all interested parties to examine, question, and support the thrust of our eorts via contributions. Here are just three reasons why YOU should consider supporting the stem cell research eorts of IAAM: 1) The present failing drug approach is a weak attempt at solving your medical problems; prescription drugs are a stop-gap measure, not a long-term, solution to critical diseases. 2) Supporting us  denitely  brings you a BIG step closer if you'd ever need stem cells. 3) Donations to IAAM are tax deductible. IAAM is a CA 501-C non-prot educational corporation. Why not rely on orthodox medicine to keep us healthy?
From an alternative, anti-aging point of view, what should be a routine protocol when visiting your doctor? Instead of just treating symptoms - - like "your cholesterol is high, I'll give you a prescription for that" - - the doctor should evaluate your health practices and guide you towards the best actions (exercise, diet, stress management, supplements, etc.) to bring out-of-range values back to normal. For example: Exercise levels  minimum for optimum results  can be precisely dened; see enclosed "Exercise, your best Ally in Aging Well (2.07)," and "Diet and Health Habits as Related to the Onset of Disease." Hans J. 1

HK Stem Cell Research - - - Facts and Fallacies

Kugler. Journal of the International Academy of Preventive Medicine, Vol.IV, No. 2, Winter 1977. Mainstream medicine's pharmaceutical drug approach has not only completely peaked out, it has actually led to a decline in our overall health - directly correlated to the increase in drug use, the overall US health rating has decreased to between 13th to 23rd place in the world, inferior even to citizens of some of the poorer nations , but it has also reached a point where medicine itself has become the # 1 cause of death in America. For details and documentation read: "Overdo$ed America: The Broken Promise of American Medicine" by John Abramson, MD, Harvard Medical School; see enclosed book review and "Death by Medicine" (8.06 Life Extension Report, full text accessible at COMMENTARY at E-books, at www.antiagingforme.com.

In Europe, stem cells (harvested from embryonic cell cultures) have been used on humans for several
years now. Scientic publications and clinical feedback from European doctors have brought out two key ndings:

a) These stem cells give the body a tremendous boost; covering a wide spectrum from immune enhancement to general revitalization. b) However, it is becoming more and more obvious that these stem cells DO NOT fully settle down in the recipient's body, forming neither new tissues nor new organ cells.

For stem cells to be truly eective, and to settle down in the recipient's body, THE STEM CELLS MUST HAVE THE RECIPIENT'S DNA. This can be done through two processes: NUCLEAR TRANSFER & PARTHENOGENESIS; both have already been demonstrated on animal models as very eective. Parthenogenesis, virgin birth, produces stem cells by triggering an egg cell to divide (without fertilization).Scientists are now working on the precise details of the recipe for nuclear transfers and parthenogenesis.

Real stem cell science is conservative! I'm a Republican, having run for oce of US Congress, I am

uncomfortable at the thought of harvesting stem cells from embryos. Scientists have learned a lot from studying such stem cells, but again, I underscore the fact that real stem cell science has already shown that this will NOT lead to eective treatments, and that making person-specic stem cells (nuclear transfers that utilize the person's own cells) is science that endures because it oers a viable means for superior treatments. Let me explain. Person-specic stem cells allow for the use of skin (or other) cells without a man and a woman, without fertilization, without growth in a womb. stem cells. Do you believe that you could go to your neighborhood hospital/clinic and ask them to make you a set of your own stem cells (with your own DNA)? Then come back a few days later and have them administered to you? Obviously not, considering the current political climate surrounding stem cells; if a need for stem cells should arise, it would be good to be connected to IAAM. IAAM stem cell research is devoted to making person-specic stem cells. To do so eciently, we need

Now, imagine for a moment that scientists had worked out all the details for making person-specic

we can hire, the faster we advance. Furthermore, biological research materials are quite expensive, and the purchase of a "Nomarski Modulation" microscope ($24,000) would accelerate nuclear transfer procedures. The speed of research is dictated exponentially according to the amount of money that is available. Please remember: IAAM is a CA 501-C non-prot corporation; donations are tax-deductible. More questions? See frequently asked Questions and Answers, below. Dr. Hans J. Kugler, PhD President, International Academy of Anti-Aging Medicine 2

your nancial support. For example: $ 50,000 can cover the cost of a research PhD for 6 months, full-time or one year part-time (we have two who have indicated interest in working with us), and $ 25,000 pays the salary of a microbiologist/microscopist, BS, for 6 months; the more scientists

HK Stem Cell Research - - - Facts and Fallacies Part II: Frequently asked Questions and Answers: Q.: Why is there an opposition to stem cells from some religious groups?
A: People who object to stem cell research for religious reasons still believe that embryos are killed in order to harvest stem cells. On the contrary, stem cell research via nuclear transfer does not involve the use of embryos. As a matter of fact, when doing nuclear transfers a patient's skin cell is manipulated  outside any womb, without any union between a man and a woman, without fertilization, and without development in a womb. Instead, nuclear transfer only requires a Petri-dish to grow patient-specic stem cells.

Q.: Any other reasons why religious people object to stem cells?

A: When people are ill-informed, they harbor worries that are unfounded. The idea that pregnancies might be induced to create a later-aborted fetus to harvest stem cells, demonstrates a lack of understanding of the science; NO STEM CELLS WITH PERSON-SPECIFIC DNA CAN EVER BE CREATED IN THIS WAY! Their worries are unsubstantiated.

Q.: Is there a way to explain why stem cell injections (harvested from embryos, DNA other than the recipient's) give the body such a tremendous boost, especially in enhancing immune functions?
A: An overall enhancement to the body as a result of stem cell injections occurs because of mechanisms similar to Dr. Paul Niehans' "Zelltherapie." Already 50 years ago, the rich and famous journeyed to Switzerland or Germany to receive embryonic sheep cell injections. The most basic growth factors, organ-specic peptides and key nucleic acids are essentially the same in humans and animals, viz., sheep, cows, and pigs; stem cells are concentrated with these powerful embryonic substances.

Q.: Is Dr. Nienhans' Zelltherapie still used in Europe?

A.: Yes, this therapy is still being used especially with children who have chromosomal abnormalities, like "Cris de Chat" (cat's cry) and Down's Syndrome. Down's syndrome children, when diagnosed and treated early enough, show a doubling of the IQ (from average 40 to 80) and facial features and height are closer to normal values according to Professor Franz Schmid, MD, Childrens Hospital, Aschaenburg, Germany. A.: No! Actually there were many renements. In one specic research project, at Heidelberg and Tuebingen universities, researchers identied many of the growth factors, low molecular weight nucleic acids and special peptides that are key factors in cellular functioning. The result of their ndings were rened into a new treatment method, "Enbryonic Cell Extracts," by Ulrich Friedrichson, MD, PhD. There are many organspecic cell extracts, ranging from mesenchym to thymus, heart, spleen, liver, eyes, brain, bone marrow, and more. Used as injections, some of the treatment results are absolutely amazing; they range from the slowing of aging, to the cessation of cancer, heart disease, and the signicant delaying of the progression of Parkinson's, to the regenerating of liver functions, reduction in herpes outbreaks, and more. For more details about cell extracts, at www.antiagingforme.com, go to COMPENDIUM and scroll down to "Embryonic Cell Extracts." Q: Who would benet from a delay in stem cell research? A: In trying to pinpoint where the resistance to stem cells may come from, pharmaceutical companies and people with nancial interests in this industry, have been on top of the list. Once stem cells become widely used all over the world, curing diseases, repairing damaged organs or entire organ-systems, and enhancing overall wellness, another industry that hinges on disease  the pharmaceutical companies  will of necessity see a downturn in prots.

Q.: Have other areas of Zelltherapie been discontinued?

Q.: What about the recent  Nov. 07  ndings by Oregon researchers re. a) nuclear transfer with monkey skin cells and b) virus-induced stem cells?

A.: Fascinating research. The making of monkey stem cells via DNA nuclear transfer from a skin cell has demonstrated that this method denitely works, and the detailed laboratory procedures outlined in this paper have brought us a big step closer to achieving our goal on human cells. The second set of publications demonstrated that, by inserting 4 viruses (each aecting dierent genes 3

HK Stem Cell Research - - - Facts and Fallacies

in the cell), a skin cell could be made to revert to the stage of a stem cell. While hailed as a new method of making stem cells, the reality is somewhat questionable; how would a stem cell with 4 viruses function when injected into a human body? Did the viruses reset the telomeres (true indicators of cell vitality) in the cell?

Q.: Are there truly conservative scientists who support nuclear transfer to make stem cells?
A.: Oh, yes! Check out "The Language of God: A Scientist Presents Evidence for Belief" by Francis Collins, MD, PhD, Free Press 2006. What a magnicent book! Every paragraph is worth reading/quoting. Previously a gene hunter at the University of Michigan at Ann Arbor, today Dr. Collins oversees a giant research empire at the National Institute of Health, the National Human Genome Research Institute in Washington. He is considered the nation's top geneticist. Here are his own words why he supports nuclear transfer to make person-specic stem cells. "There is a dierence between doing research on an embryo that was generated by sperm and egg coming together, which is the way human beings are created, versus the very bizarre laboratory phenomenon of taking a nucleus from a skin cell or the udder cell of a sheep and putting it into an environment that takes it back in time to its stem cell state."

Q.: What books do we recommend for further reading?

For the professional, with an advanced degree in a related eld, who wants to know everything about the present state of stem cell research: "Essentials of Stem Cell Biology," edited by Robert Lanza, John Gearhart, Brigid Hogan, Douglas Melton, Roger Pedersen, and many others, with 146 contributors from any possible eld(s) of stem cell research. In essence a textbook for a one-year graduate level course. Elsevier Academic Press, 546 pages, $ 129.00. The only negative: print is very small, hard to read. For the health care professional and well informed science reader: "STEM CELL NOW. From the Experiment That Shook the World to the New Politics of Life" by Christopher Scott (Stanford University lecturer and Executive Director of the Stanford University Center for Biomedical Ethics Program). Plume Publishing. This book is absolutely magnicent. A must for everybody: "The Truth About the Drug Companies: How They Deceive Us and What to Do About it," by Marcia Angell, MD (former Senior Editor, New England Journal of Medicine, now professor at Harvard Medical School). Random House Publishing.
Part III: Pictures of our stem cell lab

There are several inverted microscopes. Our star microscope  we call it "the space ship"  is a Nikon TE-2000U with Homan Modulation (to make living cells more visible under the microscope), also equipped with the nest, computerized, cell manipulation system (by Eppendorf, with the joystick control units right 4

HK Stem Cell Research - - - Facts and Fallacies

and left of the microscope).

Vibration-free tables for key microscopes  usually very expensive  can be built at a fraction of the cost by using tennis balls sitting tight in small cans (like cat food cans) to absorb vibrations (possibly caused by truck passing by the building).

There are centrifuges, magnetic stirrers, xed temperature waterbath and lots of glassware, Petri-dishes, pipettes, etc.

HK Stem Cell Research - - - Facts and Fallacies

Two key pieces of equipment are a NUAIRE CO2 Incubator and a liquid nitrogen (MVE) Dewar for storage of cells at very low temperatures.

Then there is a "Clean Room" with a NUAIRE BIOHOOD - - all aimed at working with cells and cell cultures and keeping them from getting infected.

Part IV: Parthenogenesis, Virgin-Birth. Parthenogenesis, Virgin-Birth: A way of fullling the stem cell needs of 50% of the population - - and without ethical objections.
IAAM News Relase, March 15, 2008. With much admiration for the ground-breaking work at Boston Children's Hospital (1), Kobe Univ. in Japan (2), SUMS Univ. in China (3), Univ. of Milan, Italy (4), Univ. S. Florida, Tampa (5), Heinrich Heine Univ., Germany (6), Michigan State Univ. (7), and many others: (For abstracts of works quoted above, and additional references, scroll down)

HK Stem Cell Research - - - Facts and Fallacies We believe that, on the basis of available data, the time is right for IAAM Stem Cell Research, Dr. Hans J. Kugler, PhD, director, to recommend that woman of child-bearing age give it

serious consideration to have a number of their egg cells collected and stored (by a cryogenic facility) for possible future need of stem cells. Today, when a child is born, parents have the choice of storing umbilical chord blood (containing stem cells) for future needs of the child. The same facilities could store egg cells. Even if it should turn out that egg cells are not suited for making individual-specic stem cells, these eggs are not wasted; they could be a blessing for other people and are like money in the bank. The IAAM Stem Cell Research laboratory was founded because of the many delays in stem cell research  legal, ethical, regulatory, political. Our goal: making safe and person-specic stem cells (stem cells with DNA the same as the recipient). Considering ourselves conservative, we feel uncomfortable with the concept of harvesting stem cells from aborted fetuses or from embryos from fertility clinics ( - - which will not yield person-specic stem cells anyway). We nd the Petri-dish Nuclear Transfer , and the Parthenogenesis methods acceptable, and our laboratory is set up for such work. We have been exploring parthenogenesis  virgin birth (getting an egg cell to divide without fertilization)  as a source of stem cells ; this would not require a nuclear transfer (it already contains only DNA from the donor), but it would also bypass ethical objections. Furthermore, since there - most likely - would be no ethical objections, manipulating such a cell with DNA from a (male) skin cell, it should be possible to nd ways to yield stem cells for the other half of the population.

(1) Dierentiation potential of histocompatible parthenogenetic embryonic stem cells.

Lengerke C, Kim K, Lerou P, Daley GQ Division of Pediatric Hematology/Oncology, Children's Hospital Boston, 300 Longwood Avenue, Boston, MA 02115, USA. Ann N Y Acad Sci. 2007 Jun;1106:209-18. Epub 2007 Mar 14. Embryonic stem cells (ESCs) hold unique promise for the development of cell replacement therapies, but derivation of therapeutic products from ESCs is hampered by immunological barriers. Creation of HLAtyped ESC banks, or derivation of customized ESC lines by somatic cell nuclear transfer, have been envisioned for engineering histocompatible ESC-derived products. Proof of principle experiments in the mouse have demonstrated that autologous ESCs can be obtained via nuclear transfer and dierentiated into transplantable tissues, yet nuclear transfer remains a technology with low eciency. Parthenogenesis provides an additional means for deriving ESC lines. In parthenogenesis, articial oocyte activation initiates development without sperm contribution and no viable ospring are produced in the absence of paternal gene expression. Development proceeds readily to the blastocyst stage, from which parthenogenetic ESC (pESC) lines can be derived with high eciency. We have recently shown that when pESC lines are derived from hybrid mice, early recombination events produce heterozygosity at the major histocompatibility complex (MHC) loci in some of these lines, enabling the generation of histocompatible dierentiated cells that can engraft immunocompetent MHC-matched mouse recipients. Here, we explore the dierentiation potential of murine pESCs derived in our laboratory.

(2) The maternal nucleolus is essential for early embryonic development in mammals.

Ogushi S, Palmieri C, Fulka H, Saitou M, Miyano T, Fulka J Jr., Graduate School of Science and Technology, Kobe University, Kobe 657-8501, Japan. ogushi@cdb.riken.jp Science. 2008 Feb 1;319(5863):613-6. With fertilization, the paternal and maternal contributions to the zygote are not equal. The oocyte and spermatozoon are equipped with complementary arsenals of cellular structures and molecules necessary for the creation of a developmentally competent embryo. We show that the nucleolus is exclusively of mater7

HK Stem Cell Research - - - Facts and Fallacies

nal origin. The maternal nucleolus is not necessary for oocyte maturation; however, it is necessary for the formation of pronuclear nucleoli after fertilization or parthenogenetic activation and is essential for further embryonic development. In addition, the nucleolus in the embryo produced by somatic cell nuclear transfer originates from the oocyte, demonstrating that the maternal nucleolus supports successful embryonic development.

(3) Derivation of human embryonic stem cell lines from parthenogenetic blastocysts.
Main Q, YU Y, LI T, Wang L, Chen MJ, Huang SZ, Zhou C, Zhou Q. 1Reproductive Medical Center, the First Aliated Hospital of SUMS University, Guangzhou 210029, China. Cell Res. 2007 Dec;17(12):1008-19. Parthenogenesis is one of the main, and most useful, methods to derive embryonic stem cells (ESCs), which may be an important source of histocompatible cells and tissues for cell therapy. Here we describe the derivation and characterization of two ESC lines (hPES-1 and hPES-2) from in vitro developed blastocysts following parthenogenetic activation of human oocytes. Typical ESC morphology was seen, and the expression of ESC markers was as expected for alkaline phosphatase, octamer-binding transcription factor 4, stage-specic embryonic antigen 3, stage-specic embryonic antigen 4, TRA-1-60, and TRA-1-81, and there was absence of expression of negative markers such as stage-specic embryonic antigen 1. Expression of genes specic for dierent embryonic germ layers was detected from the embryoid bodies (EBs) of both hESC lines, suggesting their dierentiation potential in vitro. However, in vivo, only hPES-1 formed teratoma consisting of all three embryonic germ layers (hPES-2 did not). Interestingly, after continuous proliferation for more than 100 passages, hPES-1 cells still maintained a normal 46 XX karyotype; hPES-2 displayed abnormalities such as chromosome translocation after long term passages. Short Tandem Repeat (STR) results demonstrated that the hPES lines were genetic matches with the egg donors, and gene imprinting data conrmed the parthenogenetic origin of these ES cells. Genome-wide SNP analysis showed a pattern typical of parthenogenesis. All of these results demonstrated the feasibility to isolate and establish human parthenogenetic ESC lines, which provides an important tool for studying epigenetic eects in ESCs as well as for future therapeutic interventions in a clinical setting.

(4) Parthenotes as a source of embryonic stem cells.

Brevini TA, Gandol F Laboratory of Biomedical Embryology, Centre for Stem Cell Research, University of Milan, Milan, Italy. tiziana.brevini@unimi.it Cell Prolif. 2008 Feb;41 Suppl 1:20-30. The derivation and study of human embryonic stem cell lines, despite their potential therapeutic usefulness, raise considerable ethical, religious, legal and political concerns because it inevitably leads to the destruction of viable embryos. In an attempt to bridge the division between ethical questions and potential scientic and medical benets, considerable eorts have been devoted to the search for alternative sources of pluripotent cell lines. In this review we discuss the use of articial parthenogenesis as a way to create entities, called parthenotes, that may represent an alternative ethical source for pluripotent cell lines. We describe the biological dierences between parthenotes and embryos, in order to provide a rationale for the discussion on whether their use can be acceptable as a source of stem cells. We present data derived from animal models on the extent parthenogenetic stem cells are similar to biparental cell lines and discuss these aspects in the context of their extension to the human species. Finally, we present experiments recently carried out in our laboratory that allowed us to generate human parthenotes through articial activation of human oocytes and to use them as a source for the derivation of parthenogenetic pluripotent cell lines.

(5) Telomere lengthening early in development.

Lui L, Bailey SM, Okuka M, Munoz P, Li C, Zhou L, Wu C, Czerwiec E, Sandler L, Seyfang A, Blasco MA, Keefe DL. Laboratory for Reproductive Medicine, Department of Obstetrics and Gynecology, University of South Florida College of Medicine, Tampa, Florida 33612, USA. liutelom@yahoo.com 8

HK Stem Cell Research - - - Facts and Fallacies

Nat.Cell Biol. 2007 Dec;9(12):1436-41. Epub 2007 Nov 4. Stem cells and cancer cells maintain telomere length mostly through telomerase. Telomerase activity is high in male germ line and stem cells, but is low or absent in mature oocytes and cleavage stage embryos, and then high again in blastocysts. How early embryos reset telomere length remains poorly understood. Here, we show that oocytes actually have shorter telomeres than somatic cells, but their telomeres lengthen remarkably during early cleavage development. Moreover, parthenogenetically activated oocytes also lengthen their telomeres, thus the capacity to elongate telomeres must reside within oocytes themselves. Notably, telomeres also elongate in the early cleavage embryos of telomerase-null mice, demonstrating that telomerase is unlikely to be responsible for the abrupt lengthening of telomeres in these cells. Coincident with telomere lengthening, extensive telomere sister-chromatid exchange (T-SCE) and colocalization of the DNA recombination proteins Rad50 and TRF1 were observed in early cleavage embryos. Both T-SCE and DNA recombination proteins decrease in blastocyst stage embryos, whereas telomerase activity increases and telomeres elongate only slowly. We suggest that telomeres lengthen during the early cleavage cycles following fertilization through a recombination-based mechanism, and that from the blastocyst stage onwards, telomerase only maintains the telomere length established by this alternative mechanism.

(6) Can articial parthenogenesis sidestep ethical pitfalls in human therapeutic cloning? An historical perspective.

Fangerau H. Institute for the History of Medicine, Heinrich-Heine University Dsseldorf, Universitaetsstrasse 1, 40225 Dsseldorf, Germany. heiner.Fangerau@uni-duesseldorf.de J Med Ethics. 2005 Dec;31(12):733-5. The aim of regenerative medicine is to reconstruct tissue that has been lost or pathologically altered. Therapeutic cloning seems to oer a method of achieving this aim; however, the ethical debate surrounding human therapeutic cloning is highly controversial. Articial parthenogenesis-obtaining embryos from unfertilized eggs-seems to oer a way to sidestep these ethical pitfalls. Jacques Loeb (1859-1924), the founding father of articial parthenogenesis, faced negative public opinion when he published his research in 1899. His research, the public's response to his ndings, and his ethical foundations serve as an historical argument both for the communication of science and compromise in biological research.

7) Embryonic stem cells from parthenotes. Cibelli JB, Cunni K, Vrana KE.

Department of Animal Science, Michigan State University, East Lansing, Michigan, USA. Methods Enzymol. 2006;418:117-35. While human embryonic stem cells (hESCs) hold tremendous therapeutic potential, they also create societal and ethical dilemmas. Adult and placental stem cells represent two alternatives to the hESC, but may have technical limitations. An additional alternative is the stem cell derived from parthenogenesis. Parthenogenesis is a reproductive mechanism that is common in lower organisms and produces a live birth from an oocyte activated in the absence of sperm. However, parthenogenetic embryos will develop to the blastocyst stage and so can serve as a source of embryonic stem cells. Parthenogenetic ESCs (pESCs) have been shown to have the properties of self-renewal and the capacity to generate cell derivatives from the three germ layers, conrmed by contributions to chimeric animals and/or teratoma formation when injected into SCID mice. Therefore, this mechanism for generating stem cells has the ethical advantage of not involving the destruction of viable embryos. Moreover, the cells do not involve the union of male and female and so genetic material will be derived exclusively from the female oocyte donor (with the attendant potential immunological advantages). This chapter describes the biology underlying parthenogenesis, as well as provides detailed technical considerations for the production of pESCs.

Wasserman D. University of Maryland, USA. Am J Bioeth. 2005 Nov-Dec;5(6):8-16.

(8) Comment on: What qualies as a live embryo?

HK Stem Cell Research - - - Facts and Fallacies (9) Dierentiation potential of parthenogenetic embryonic stem cells is improved by nuclear transfer. Hikichi T, Wakayama S, Mizutani E, Takashima Y, Kishigami S, Van Thuan N, Ohta H, Thuy Bui H, Nishikawa S, Wakayama T.
Laboratory for Genomic Reprogramming, Center for Developmental Biology, RIKEN Kobe, Kobe, Japan. Stem Cells. 2007 Jan;25(1):46-53. Epub 2006 Sep 28.

(10) Histocompatible embryonic stem cells by parthenogenesis. Kim K, Lerou P, Yabuuchi A, Lengerke C, Ng K, West J, Kirby A, Daly MJ, Daley GQ.

Division of Pediatric Hematology/Oncology, Children's Hospital Boston and Dana Farber Cancer Institute, Boston, MA 02115, USA. Science. 2007 Jan 26;315(5811):482-6. Epub 2006 Dec 14.

(11) Ethical questions concerning research on human embryos, embryonic stem cells and chimeras.
Bobbert M. Department of Medical Ethics at the Institute for the History of Medicine, Medical Faculty of the University of Heidelberg, Heidelberg, Germany. Biotechnol J. 2006 Dec;1(12):1352-69.

Talbot NC, Powell AM, Camp M, Ealy AD. USDA, ARS, ANRI, Biotechnology and Germplasm Laboratory, Beltsville, MD 20705, USA. ntalbot@anri.barc.usda.gov In Vitro Cell Dev Biol Anim. 2007 Feb;43(2):59-71. Epub 2007 Mar 21.

(12) Establishment of a bovine blastocyst-derived cell line collection for the comparative analysis of embryos created in vivo and by in vitro fertilization, somatic cell nuclear transfer, or parthenogenetic activation.

(13) 'Virgin birth' stem cells bypass ethical objections.

Geddes L. New Sci. 2006 Jul 1-7;191(2558):19.

(14) Human leukocyte antigen matching estimations in a hypothetical bank of human embryonic stem cell lines in the Japanese population for use in cell transplantation therapy. Nakajima F, Tokunaga K, Nakatsuji N.

Research and Development Department, Central Blood Institute, Japanese Red Cross Society, Tokyo, Japan. Stem Cells 2007, Apr;25(4):983-5. Epub 2006 Dec 21. Green RM. Ethics Institute, Dartmouth College, 27 North Main Street, Hanover, New Hampshire 03755-3500, USA. ronald.m.green@dartmouth.edu Nat Rev Genet 2007 Jun;8(6):480-5.

(15) Can we develop ethically universal embryonic stem-cell lines?

(16) Birth of cloned pigs from zona-free nuclear transfer blastocysts developed in vitro before transfer.
Lagutina I, Lazarri G, Galli C. Laboratory of Reproductive Technologies, Italian Experimental Institute Lazzaro Spallanzani, Cremona, Italy. Cloning Stem Cells. Winter;8(4):283-93. Surani MA. Wellcome Trust Cancer Research UK Gurdon Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK. a.surani@gurdon.cam.ac.uk C R Biol. 2007 Jun-Jul;330(6-7):474-8. Epub 2007 May 9. 10

(17) Germ cells: the eternal link between generations.

HK Stem Cell Research - - - Facts and Fallacies (18) Characterization and multilineage dierentiation of embryonic stem cells derived from a bualo parthenogenetic embryo.
Kitiyanant Y, Kupradinum P, Mitalipov S, Kusamran T. Department of Biochemistry, Faculty of science, Mahidol University, Bangkok, Thailand. Mol Reprod Dev. 2007 Oct;74(10):1295-302.

(19)Establishment of autologous embryonic stem cells derived from preantral follicle culture and oocyte parthenogenesis.
Lee ST, Choi MH, Lee EJ, Gong SP, Jang M, Park SH, Jee H, Kim DY, Han JY, Lim JM. Gamete and Stem Cell Biotechnology Laboratory, Department of Agricultural Biology. Fertil Steril. 2007 Apr 18 [Epub ahead of print]

(20) Hematopoietic reconstitution with androgenetic and gynogenetic stem cells. Eckardt S, Leu NA, Bradley HL, Kato H, Bunting KD, McLaughlin KJ. Center for Animal Transgenesis and Germ Cell Research, New Bolton Center, University of Pennsylvania, Kennett Square, Pennsylvania 19348, USA. Genes Dev. 2007 Feb 15;21(4):409-19. (21) Towards the generation of patient-specic pluripotent stem cells for combined gene and
cell therapy of hematologic disorders. Daley GQ. Hematology Am Soc Hematol Educ Program. 2007;2007:17-22

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