Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antibodies, Monoclonal / pharmacology
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Antigens, Ly / analysis
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Bone Marrow Cells*
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Cell Adhesion Molecules / immunology
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Cell Adhesion Molecules / physiology
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Cell Separation*
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Cells, Cultured
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Flow Cytometry
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Granulocytes / cytology*
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Hematopoietic Stem Cells / chemistry
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Hematopoietic Stem Cells / cytology*
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Lymphocytes / cytology*
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Membrane Proteins / analysis
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Mice
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Mice, Inbred C57BL
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Proto-Oncogene Proteins / analysis
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Proto-Oncogene Proteins / antagonists & inhibitors
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Proto-Oncogene Proteins / physiology
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Proto-Oncogene Proteins c-kit
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Receptor Protein-Tyrosine Kinases / analysis
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Receptor Protein-Tyrosine Kinases / antagonists & inhibitors
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Receptor Protein-Tyrosine Kinases / physiology
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Receptors, Colony-Stimulating Factor / analysis
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Receptors, Colony-Stimulating Factor / antagonists & inhibitors
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Receptors, Colony-Stimulating Factor / physiology
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Receptors, Very Late Antigen / antagonists & inhibitors
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Receptors, Very Late Antigen / physiology
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Stromal Cells / physiology*
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Vascular Cell Adhesion Molecule-1
Substances
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Antibodies, Monoclonal
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Antigens, Ly
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Cell Adhesion Molecules
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Ly6a protein, mouse
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Membrane Proteins
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Proto-Oncogene Proteins
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Receptors, Colony-Stimulating Factor
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Receptors, Very Late Antigen
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Vascular Cell Adhesion Molecule-1
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Proto-Oncogene Proteins c-kit
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Receptor Protein-Tyrosine Kinases