Detection of murine adult bone marrow stroma‐initiating cells in Lin⁻c‐fms⁺c‐kit^lowVCAM‐1⁺ cells

Abstract

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.

Interleukin-6 deficiency affects bone marrow stromal precursors, resulting in defective hematopoietic support

“…Our analysis of membrane proteins in wild-type and IL-6 Ϫ/Ϫ stromal cultures showed increased expression in wild-type stroma of VCAM-1 and Sca-1, considered mesenchymal cell markers. 35,36 VCAM-1 expression has been linked to the hematopoietic supportive capacity of immortalized murine BM stromal cell lines. 29 Sca-1 expression was also correlated with the ability of a murine BM stromal cell line to support proliferation of 5-fluorouracil (5-FU)-treated BM cells.…”

Myeloid-specific expression of Stat3C results in conversion of bone marrow mesenchymal stem cells into alveolar type II epithelial cells in the lung

“…Modification of the glycans on CD44 increases the efficiency of BMSCs delivery and homing to the bone marrow [ 31 ]. CD106 expression has been linked to the hematopoietic supportive capacity of immortalized murine BM stromal cell lines [ 38 , 39 ]. In our analysis, expression of Api6 was also high in cultured BMSCs.…”