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Hematopoietic Stem Cells

Identification of Hematopoietic Stem Cells in Human and Mouse

The first isolation of hematopoietic stem cells (HSC) required quantitative clonal assays for every blood cell progenitor type (T, B, myeloerythroid) and methods to sort cells based on their unique expression profiles, as determined by cell surface markers. Using this approach, both mouse and human hematopoietic stem cells were prospectively isolated. All HSC activity in adult mouse bone marrow (BM) was shown to be contained in a population marked by the composite phenotype of c-Kit+, Thy-1.1lo, lineage marker-, and Sca-1+ (designated KTLS). These KTLS hematopoietic stem cells in mice transplanted at the single-cell level gave rise to lifelong hematopoiesis, including a steady state of 20 to 100,000 HSC and over 109 blood cells produced daily.

Mouse Hematopoietic Stem Cell Identification

Mouse Hematopoietic Stem Cell Identification

Mouse HSC Identification

C57Bl/6 bone marrow cells were stained with the Mouse Hematopoietic Lineage Flow Cocktail, APC Anti-Mouse CD117 and PE Anti-Mouse Sca-1. Viable cells in the lymphocyte gate were used for analysis. Events displayed on the right plot are gated on lineage negative events, as shown on the left histogram.

Human Hematopoietic Stem Cell Identification

In humans, the isolation of a lin- CD34+CD90+ progenitor cell resulted in the purification of a homogeneous HSC population. The critical test for HSCs is in their ability to clinically rescue myelo-ablated hosts from hematopoietic failure, and establish long-term multi-hematopoietic lineage reconstitution. Clinical implantation of HSCs into cancer patients has led to stable grafts depleted of contaminating hematopoietic cells and resident or metastasized malignant cells. The continued identification of novel hematopoietic stem cell markers will enable improved efficiency in the purification of HSCs for the treatment of disease.

Human Hematopoietic Stem Cell Identification

Human HSC Identification

Normal human peripheral blood cells were stained with the FITC Human Hematopoietic Lineage Flow Cocktail, Alexa Fluor® 647 Anti-Human CD34 and PE Anti-Human CD59. Total viable cells were used for analysis. Cells in the right dot plot are gated on lineage negative events, depicted in the left histogram.

Phenotypes of Hematopoietic Stem Cells

The tables below outline the accepted phenotypes for the isolation of human and mouse hematopoietic stem cells and developmental intermediates of the lymphoid and myeloid lineages.

Mouse Hematopoietic Stem Cell Identification Schemes
NamePhenotypeOriginLocation
Multipotent, CD34/Flk2 Series      
LT.HSC.34F.BM CD34-Flk2-Lin-c-Kit+Sca1+ B6 BM
ST.HSC.34F.BM CD34+Flk2-Lin-c-Kit+Sca1+ B6 BM
MPP.34F.BM CD34+Flk2+Lin-c-Kit+Sca1+ B6 BM
Multipotent, SLAM Series      
LT.HSC.SL.BM CD150+CD48-Lin-c-Kit+Sca1+ B6 BM
ST.HSC.SL.BM CD150-CD48-Lin-c-Kit+Sca1+ B6 BM
Oligopotent      
CLP.BM Flk2+IL7Rα+c-Kit+Sca1+Lin- B6 BM
CMP.BM Lin-Thy1-IL7Rα-Sca1-c-Kit+FcγRIloCD34+ B6 BM
MEP.BM Lin-Thy1-IL7Rα-Sca1-c-Kit+FcγRIloCD34- B6 BM
GMP.BM Lin-Thy1-IL7Rα-Sca1-c-Kit+FcγRIhiCD34+ B6 BM
Human Hematopoietic Stem Cell Identification Schemes
NamePhenotype
Hematopoietic Stem Cell Lin-CD34+CD38-CD90+CD45RA-
Multipotent Progenitor Lin-CD34+CD38-CD90-CD45RA-
Common Myeloid Progenitor Lin-CD34+CD38+IL-3RαloCD45RA-
Common Lymphoid Progenitor Lin-CD34+CD38+CD10+

 

References

The promise of human induced pluripotent stem cells for research and therapy. Nishikawa S, Goldstein RA, Nierras CR. Nat Rev Mol Cell Biol. 2008 Sep;9(9):725-9. Review.