Identification of a population of cells with hematopoietic stem cell properties in mouse aorta-gonad-mesonephros cultures.
The aorta-gonad-mesonephros (AGM) region is a primary source of definitive hematopoietic cells in the midgestation mouse embryo. In cultures of dispersed AGM regions, adherent cells containing endothelial cells are observed first, and then non-adherent hematopoietic cells are produced. Here we report on the characterization of hematopoietic cells that emerge in the AGM culture. Based on the expression profiles of CD45 and c-Kit, we defined three cell populations: CD45(low) c-Kit(+) cells that had the ability to form hematopoietic cell colonies in methylcellulose media and in co-cultures with stromal cells; CD45(low) c-Kit(-) cells that showed a granulocyte morphology; CD45(high) c-Kit(low/-) that exhibited a macrophage morphology. In co-cultures of OP9 stromal cells and freshly prepared AGM cultures, CD45(low) c-Kit(+) cells from the AGM culture had the abilities to reproduce CD45(low) c-Kit(+) cells and differentiate into CD45(low) c-Kit(-) and CD45(high) c-Kit(low/-) cells, whereas CD45(low) c-Kit(-) and CD45(high) c-Kit(low/-) did not produce CD45(low) c-Kit(+) cells. Furthermore, CD45(low) c-Kit(+) cells displayed a long-term repopulating activity in adult hematopoietic tissue when transplanted into the liver of irradiated newborn mice. These results indicate that CD45(low) c-Kit(+) cells from the AGM culture have the potential to reconstitute multi-lineage hematopoietic cells.
Nobuhisa
Ikuo
I
Division of Cell Fate Modulation, Institute of Molecular Embryology and Genetics, Kumamoto University, 2-2-1 Honjo, Kumamoto, Japan.
Ohtsu
Naoki
N
Okada
Seiji
S
Nakagata
Naomi
N
Taga
Tetsuya
T
D000818:Animals; D001011:Aorta / Q000166:cytology* / Q000196:embryology; D002454:Cell Differentiation; D016207:Cytokines / Q000494:pharmacology; D006066:Gonads / Q000166:cytology* / Q000196:embryology; D006412:Hematopoietic Stem Cells / Q000378:metabolism / Q000502:physiology*; D017493:Leukocyte Common Antigens / Q000378:metabolism; D008650:Mesonephros / Q000166:cytology* / Q000196:embryology; D051379:Mice; D008813:Mice, Inbred ICR; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D033581:Stem Cell Transplantation; D017154:Stromal Cells / Q000502:physiology; D046509:Tissue Culture Techniques / Q000379:methods*
D016207:Cytokines; D019009:Proto-Oncogene Proteins c-kit; D017493:Leukocyte Common Antigens