Prospective isolation and molecular characterization of hematopoietic stem cells with durable self-renewal potential.
Hematopoietic stem cells (HSCs) are generally defined by their dual properties of pluripotency and extensive self-renewal capacity. However, a lack of experimental clarity as to what constitutes extensive self-renewal capacity coupled with an absence of methods to prospectively isolate long-term repopulating cells with defined self-renewal activities has made it difficult to identify the essential components of the self-renewal machinery and investigate their regulation. We now show that cells capable of repopulating irradiated congenic hosts for 4 months and producing clones of cells that can be serially transplanted are selectively and highly enriched in the CD150(+) subset of the EPCR(+)CD48(-)CD45(+) fraction of mouse fetal liver and adult bone marrow cells. In contrast, cells that repopulate primary hosts for the same period but show more limited self-renewal activity are enriched in the CD150(-) subset. Comparative transcriptome analyses of these 2 subsets with each other and with HSCs whose self-renewal activity has been rapidly extinguished in vitro revealed 3 new genes (VWF, Rhob, Pld3) whose elevated expression is a consistent and selective feature of the long-term repopulating cells with durable self-renewal capacity. These findings establish the identity of a phenotypically and molecularly distinct class of pluripotent hematopoietic cells with lifelong self-renewal capacity.
Kent
David G
DG
Terry Fox Laboratory, British Columbia Cancer Agency, Vancouver, BC, Canada.
Copley
Michael R
MR
Benz
Claudia
C
Wöhrer
Stefan
S
Dykstra
Brad J
BJ
Ma
Elaine
E
Cheyne
John
J
Zhao
Yongjun
Y
Bowie
Michelle B
MB
Zhao
Yun
Y
Gasparetto
Maura
M
Delaney
Allen
A
Smith
Clayton
C
Marra
Marco
M
Eaves
Connie J
CJ
D000818:Animals; D020296:Animals, Congenic; D015703:Antigens, CD / Q000032:analysis; D000943:Antigens, Differentiation / Q000032:analysis; D001854:Bone Marrow Cells / Q000166:cytology; D002455:Cell Division; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured / Q000637:transplantation; D005434:Flow Cytometry / Q000379:methods*; D020869:Gene Expression Profiling; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D016130:Immunophenotyping; D017493:Leukocyte Common Antigens / Q000032:analysis; D008099:Liver / Q000166:cytology / Q000196:embryology; D051379:Mice; D008810:Mice, Inbred C57BL; D010739:Phospholipase D / Q000032:analysis; D011828:Radiation Chimera; D011956:Receptors, Cell Surface / Q000032:analysis; D000071177:Signaling Lymphocytic Activation Molecule Family Member 1; D020743:rhoB GTP-Binding Protein / Q000032:analysis / Q000235:genetics; D014841:von Willebrand Factor / Q000032:analysis / Q000235:genetics; D005090:Exodeoxyribonucleases
D015703:Antigens, CD; D000943:Antigens, Differentiation; D017493:Leukocyte Common Antigens; D010739:Phospholipase D; D011956:Receptors, Cell Surface; D000071177:Signaling Lymphocytic Activation Molecule Family Member 1; D020743:rhoB GTP-Binding Protein; D014841:von Willebrand Factor; C516770:Ptprc protein, mouse; C542157:Pld3 protein, mouse; D005090:Exodeoxyribonucleases