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(HSC).
HSCs are pluripotent (can give rise to differentiated blood cells of all lineages T & B & NK lymphocytes, erythrocytes, mast cells, megakaryocytes & platelets, eosinophils, basophils, neutrophils, monocytes/macrophages , etc.). Lack the markers of differentiated blood
HSCs are rare, mainly quiescent, undifferentiated cells that on occasion produce by mitosis 2 kinds of progeny: more stem cells (self-renewal) cells that begin to differentiate along the various paths to terminally differentiated hematopoietic lineages
Path of differentiation is generally regulated by the need for more of that particular type of blood cell, and is controlled by the appropriate cytokines and hormones and growth factors, colony stimulating factors (CSFs)
lymphocytes terminally diff.: can undergo further division (i.e. memory cells)
anuclear
Lineage diagram outline of how hematopoietic cells are increasingly restricted in the types of progeny to which they can give rise as differentiation proceeds
For many years the YS was assumed to be the primary site of formation of the HSCs that migrate to and colonize the fetal liver and subsequently the bone marrow.
d. 31-34
1st DEFINITIVE multipotent hematopoietic stem cells are generated within the embryonic AGM region of the para-aortic splanchnopleuric mesoderm
(d. 30-37 human, and late d. 10/early d. 11 mice) DEFINITIVE HEMATOPOIESIS HSCs can restore long-term multilineage hematopoiesis when transplanted into adult myeloablated recipients, and generates enucleated erythrocytes, various kinds of myeloid and lymphoid cells, and long-term reconstituting hematopoietic stem cells (LTR-HSCs) Upon transplantation into NODSCID mice, cultured AGM HSC cells showed lymphomyeloid reconstitution; YS cells were only capable of contributing to the myeloid lineage (Tavian et al., 2001).
The fetal liver is the main organ for hematopoiesis during fetal life
After about d. 37 (5 weeks), HSCs from the AGM begin to colonize the fetal liver, and at 6 weeks hematopoiesis (definitive) takes place in the fetal liver until bone marrow is formed Throughout fetal life, the liver is the chief organ for production of myeloid and erythroid cells.
Bone marrow (BM) is seeded by liver HSCs by 8 weeks. B-lymphopoiesis takes place in liver @ 7 wks., then shifts to bone marrow.
Hematopoiesis from fetal BM is mainly myeloid and contributes only minimally to the blood pool throughout fetal life. After birth, BM becomes main hematopoietic organ. Many HPC and HSC in circulation during fetal life and immediately after birth; 24-48 hours after birth they disappear due to lodgement in BM.
Bone marrow: site of pre- and peri-natal, childhood, and adult hematopoiesis
During fetal growth, hematopoiesis takes place in all bony cavities (axial and appendicular skeleton) as well as in liver and spleen. Prior to birth, splenic and hepatic hematopoiesis disappear, and gradually thereafter hematopoietic tissue (red marrow) is replaced by adipocytes (yellow marrow) beginning in the distal bones and retracting to the adult pattern by age ten. Yellow marrow can be reactivated by an increased demand for blood cells, i.e., blood loss, but does not normally produce blood cells In the adult, hematopoietic marrow is confined to the axial skeleton (sternum, vertebrae, iliac bones, ribs) and proximal portions of the humerus and femur.
Bone marrow is specially construed to support the proliferation, differentiation, and maintenance of hematopoietic cells
Honeycombed latticework of venous sinuses large, thin-walled veins Endothelial cells lining the marrow sinuses are bounded by STROMAL CELLS that generate an EXTRACELLULAR MATRIX that mechanically support hematopoietic cells and vasculature; provide a nurturing microenvironment for hematopoiesis & hematopoietic colonies Adjacent cells in marrow: endothelial cells, fibroblasts, adipocytes, osteoblasts, and macrophages and reticular connective tissue. Close contact between hematopoietic cells and these cells, especially the stroma facilitates transmission of proliferative signals or diffusion of locally produced cytokines Maturing blood cells can enter the circulation through openings in the vascular sinuses (megakaryocytes & erythroblasts clustered against sinuses) usually go first to other hematopoietic tissues for further maturation
hematopoietic stem cell (HSC). HSCs are pluripotent (can give rise to differentiated blood cells of all lineages T & B & NK lymphocytes, erythrocytes, mast cells, megakaryocytes & platelets, eosinophils, basophils, neutrophils, monocytes/macrophages, DCs, etc.)
HSCs are rare (1 in every 105 nucleated cells in adult bone marrow)
Are mainly quiescent, undifferentiated cells that on occasion produce by mitosis 2 kinds of progeny (asymmetric division) :
more stem cells (HSCs have a limited ability to self-renew) progenitor cells that can undergo further divisions and become progressively more differentiated and more restricted in their capacity for self renewal
Functional assay to ID stem cells in vivo: is the cell able to stably generate multiple hematopoietic lineages in irradiated or SCID (immunodeficient) mice?
Have done this with human HSCs, termed SCID repopulating cells or SRCs able to repopulate human hematopoiesis in immunodeficient mice -- LTR long-term repopulation SRC homes to & engrafts murine BM and produces immature cells Capable of multilineage difftn Only found in CD34+/38- frxn -- purify these to get enriched HSC population, and/or select for VEGF receptor expression After cells are transplanted, a CD34+/CD38- population of cells supports short term hematopoiesis, following which a CD34-/CD38- population supports long-term reconstitution
48 hrs
XX
Short-term reconstituting XX blood cells XY Serial transplantation to 2ndary host
XX
Blood is XY
Progenitor cells are multipotent do not self-renew or have only an extremely limited capacity respond best to multiple cytokines is a compartment of hematopoiesis that expands the number of cells dramatically are named by the types of colonies they give rise to:
The pluripotent HSC gives rise to lymphoid and myeloid stem cells, the latter of which gives rise to a GEMM progenitor termed CFU-GEMM
CFU-GEMM is a multipotent cell giving rise to granulocyte, erythroid, monocyte, and megakaryocyte colonies CFU-GM gives rise to both granulocyte and monocyte colonies
Cell cycle status is tightly associated with the function of cells at each step of hematopoiesis: 1) Primitive stem cells (CD34+) slow cell cycling or dormancy 2) Progenitor populations rapid cycling required for effective expansion 3) Terminally differentiated cells withdrawal from cell cycle suitable and sometimes prerequisite for functions of mature cells
Cell cycle status is tightly associated with the function of cells at each step of hematopoiesis: 1) Primitive stem cells (CD34+) slow cell cycling or dormancy 2) Progenitor populations rapid cycling required for effective expansion 3) Terminally differentiated cells withdrawal from cell cycle suitable and sometimes prerequisite for functions of mature cells
Differentiation and lineage commitment occur under the influence of a complex array of signals from the extracellular environment, especially cytokines such as stem cell factor (c-Kit ligand), IL-3, GM-CSF, and G-CSF binding of cytokines to cell surface receptors results in the initiation of a cascade of signal transduction events within the cell.
Series of negative regulators & amplification circuits provide additional control over the process of hematopoiesis.
Neutrophils
Major component of innate immune system 1st line of defense against infection
A key concept : The marrow contains a large storage pool of neutrophils which can be reserved for release in a setting of stress, and that the exponential expansion of progenitor cells can be augmented by granulocyte colony stimulating factor (G-CSF) under stress conditions. Neutrophils and Host Defense Most are in the marrow--reserve Circulating neutrophils: half in vessel, half adherent to wall Have peptide receptors for and diapedese in response to FMLP (Nformyl-methionyl-leucyl-phenylalanine), chemotaxins Steady flow of neutrophils into superficial tissue, skin, mucosa, lungs needed to prevent infection. Bactericidal mechanisms Degradative enzymes in granules Oxidative killing Superoxide generation by NADPH-dependent oxidase Involves hexose monophosphate shunt, cytochrome b Patients with Chronic Granulomatous Disease lack cytochrome b cannot generate superoxide, and develop repeated infections.
Example of differentiation from the precursor stage onward: myeloblast to granulocyte (neutrophil maturation)
billion cells per kg
0.51 1.95
0.14
2.7
myeloblast
myelocyte
po
3.6
metamyelocyte
2.5
band neutrophil
Increasing phagocytic function pseudopodia extend around microorganisms and fuse to form a phagosome into which granuole contents are released
neutrophil
Expansion of cell number occurs as cells in the mitotic or proliferative pool replicate
The post-mitotic pool can no longer divide but continues to mature into terminally differentiated cells
In the setting of infection or stress, maturation time may be shortened, divisions may be skipped, and cells may be released into the bloodstream earlier
general adhesion molecules are expressed on hematopoietic progenitor cells in the marrow but are downregulated or degraded to facilitate egress of progenitors from the BM -- selectins, selectin ligands, integrins, CD44 and PECAM
Mobilized PBL cells have a much faster engraftment than do bone marrow cells, due to the increased cell dose of transplanted mobilized cells and increased numbers of committed progenitor cells. BUT proliferation & differentiation potential of CD34+/CD38- stem cells from mobilized PBL is inferior to that of undifferentiated BM.
Cord blood stem cells: Immediately after birth, relatively high levels of immature CD34 + progenitor & stem cells circulating (for about 48 hours) umbilical cord blood represents a promising source of stem cells for transplantation, but #s are too low for transplant into adults need to find a way to expand ex vivo (stromal cells).
Model of mechanisms of stem cell mobilization by G-CSF: disruption of retention in BM & proximity to stromal cells
1) Disruption of adhesion interactions btwn. HSC and BM stromal microenvironment VLA-4 is an integrin on HSCs that binds to VCAM-1 (another adhesion molecule) on stromal cells -- needs to be downregulated for HSCs to egress. HSCs also increase the expression of proteolytic enzymes elastase MMPs, and cathepsin G during mobilization so that they can cleave VCAM-1 from the stromal cells. 2) Disruption of chemotactic interactions SDF-1 retention signal produced by BM stromal cells gets degraded by same proteolytic enzymes CXCR4 is upregulated on HSCs in mobilization why? Probably important for egress to interact with SDF-1 in the blood.