Bone marrow stromal cells induce myeloid and lymphoid development of the sorted hematopoietic stem cells in vitro.
Regulation of development of hematopoietic stem cells was examined by culturing Lin- c-Kit+ Sca1+ stem cells sorted from bone marrow (BM) cells by fluorescence-activated cell sorting on a layer of TBR59, a BM stromal cell line established from simian virus 40 T-antigen gene transgenic mice. The sorted stem cells did not show self-renewal, but two waves (at 7 and 13 days) of a cobblestone formation were induced by the stromal cell layer. The cobblestones were formed by finite cell division (eight divisions on average) of sorted Lin- c-Kit+ Sca1+ stem cells, and divided cells were still immature. The c-Kithigh stem cell population was induced to form the first wave of cobblestone formation committed to myeloid lineage, whereas c-Kitlow population was induced to form the second wave of this formation committed to lymphoid lineage. Both cobblestone formations require c-Kit function, but very late activation antigen-4-vascular cell adhesion molecule-1 interaction plays different parts in the two lineages.
Okuyama
R
R
Department of Cell Biology, Tohoku University, Sendai, Japan.
Koguma
M
M
Yanai
N
N
Obinata
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D000950:Antigens, Ly / Q000032:analysis; D001854:Bone Marrow Cells; D015815:Cell Adhesion Molecules / Q000276:immunology / Q000502:physiology; D002469:Cell Separation; D002478:Cells, Cultured; D005434:Flow Cytometry; D006098:Granulocytes / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000166:cytology*; D008214:Lymphocytes / Q000166:cytology*; D008565:Membrane Proteins / Q000032:analysis; D051379:Mice; D008810:Mice, Inbred C57BL; D011518:Proto-Oncogene Proteins / Q000032:analysis / Q000037:antagonists & inhibitors / Q000502:physiology; D019009:Proto-Oncogene Proteins c-kit; D020794:Receptor Protein-Tyrosine Kinases / Q000032:analysis / Q000037:antagonists & inhibitors / Q000502:physiology; D016184:Receptors, Colony-Stimulating Factor / Q000032:analysis / Q000037:antagonists & inhibitors / Q000502:physiology; D016029:Receptors, Very Late Antigen / Q000037:antagonists & inhibitors / Q000502:physiology; D017154:Stromal Cells / Q000502:physiology*; D019010:Vascular Cell Adhesion Molecule-1
D000911:Antibodies, Monoclonal; D000950:Antigens, Ly; D015815:Cell Adhesion Molecules; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D011518:Proto-Oncogene Proteins; D016184:Receptors, Colony-Stimulating Factor; D016029:Receptors, Very Late Antigen; D019010:Vascular Cell Adhesion Molecule-1; D019009:Proto-Oncogene Proteins c-kit; D020794:Receptor Protein-Tyrosine Kinases