Detection of murine adult bone marrow stroma-initiating cells in Lin(-)c-fms(+)c-kit(low)VCAM-1(+) cells.
We attempted to characterize the phenotype of cells which initiate fibroblastic stromal cell formation (stroma-initiating cells: SICs), precursor cells for fibroblastic stromal cells, based on the expression of cell surface antigens. First, we stained adult murine bone marrow cells with several monoclonal antibodies and separated them by magnetic cell sorting. SICs were abundant in the c-kit(+), Sca-1(+), CD34(+), VCAM-1(+), c-fms(+), and Mac-1(-) populations. SICs were recovered in the lineage-negative (Lin(-)) cells but not the Lin(+) cells. When macrophage colony-stimulating factor (M-CSF) was absent from the culture medium, no stromal colony appeared among the populations enriched in SICs. Based on these findings, the cells negative for lineage markers and positive for c-fms (M-CSF receptor) were further divided on the basis of the expression of c-kit, VCAM-1, Sca-1 or CD34 with a fluorescence-activated cell sorter. SICs were found to be enriched in the Lin(-)c-fms(+)c-kit(low) cells and Lin(-)c-fms(+)VCAM-1(+) cells but not in Lin(-)c-fms(+)Sca-1(+) cells and Lin(-)c-fms(+)CD34(low) cells. As a result, the SICs were found to be present at highest frequency in Lin(-)c-fms(+)c-kit(low)VCAM-1(+) cells: a mean of 64% of the SICs in the Lin(-) cells were recovered in the population. In morphology and several characteristics, the stromal cells derived from Lin(-)c-fms(+)c-kit(low)VCAM-1(+) cells resembled fibroblastic cells. The number of Lin(-)c-fms(+)c-kit(low)VCAM-1(+) cells in bone marrow of mice injected with M-CSF was higher than that in control mice. In this study, we identified SICs as Lin(-)c-fms(+)c-kit(low)VCAM-1(+) cells and demonstrated that M-CSF had the ability to increase the cell population in vivo.
Tanaka-Douzono
M
M
Biochemical Research Laboratory, Morinaga Milk Industry Co. Ltd., Zama, Kanagawa, Japan.
Suzu
S
S
Yamada
M
M
Wakimoto
N
N
Hayasawa
H
H
Hatake
K
K
Motoyoshi
K
K
D000818:Animals; D000943:Antigens, Differentiation / Q000032:analysis; D001854:Bone Marrow Cells / Q000166:cytology*; D002455:Cell Division; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D005347:Fibroblasts / Q000166:cytology; D006412:Hematopoietic Stem Cells / Q000166:cytology; D018189:Immunomagnetic Separation; D016130:Immunophenotyping; D016173:Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D019009:Proto-Oncogene Proteins c-kit / Q000032:analysis* / Q000276:immunology; D016186:Receptor, Macrophage Colony-Stimulating Factor / Q000032:analysis* / Q000276:immunology; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects; D017154:Stromal Cells / Q000502:physiology*; D019010:Vascular Cell Adhesion Molecule-1 / Q000032:analysis* / Q000276:immunology
D000943:Antigens, Differentiation; D019010:Vascular Cell Adhesion Molecule-1; D016173:Macrophage Colony-Stimulating Factor; D019009:Proto-Oncogene Proteins c-kit; D016186:Receptor, Macrophage Colony-Stimulating Factor