PuSH - Publication Server of Helmholtz Zentrum München: Characterization of hemopoietic cell populations from human cord blood expressing c-kit.

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Reisbach, G. ; Bartke, I. ; Kempkes, B. ; Kostka, G. ; Ellwart, J.W. ; Birner, A. ; Thalmeier, K. ; Mailhammer, R. ; Bornkamm, G.W. ; Ullrich, A. ; Dörmér, P.G.

Characterization of hemopoietic cell populations from human cord blood expressing c-kit.

Exp. Hematol. 21, 74-79 (1993)

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Human cord blood or bone marrow cells expressing the CD34 surface antigen include a population of pluripotent progenitors. We identified and isolated a subpopulation of cells coexpressing CD34 and c-kit, a transmembrane receptor with tyrosine kinase activity. Novel monoclonal antibodies (16A6, 14A3, 3D6) directed against the extracellular domain of c-kit were used for immunofluorescence labeling and sorting of low-density mononuclear cells (MNCs) from umbilical cord blood and bone marrow. The frequency of c-kit- labeled MNCs from cord blood (mean 5.0% ± 2.1%, n=16) was similar to that from adult bone marrow (mean 3.7% ± 1.3%, n=4). On average, 1.4% of CD34- positive cells were recorded in cord blood and 2.1% in bone marrow MNCs. Roughly 60% of CD34-positive cells coexpressed c-kit. The ability of CD34+/c- kit+ cells to form multilineage colonies (CFU-GEMM) was assayed after sorting with an antibody that did not show any significant effect on c-kit ligand (RL) or granulocyte/macrophage colony-stimulating factor (GM-CSF)- induced colony formation. For CD34+/c-kit+ cells, we found a 20-to 50-fold enrichment as against total MNCs, and a 2-fold enrichment if compared with the CD34+/c-kit- population. To study expression of c-kit in lymphocytic precursors, monoclonal anti-CD7 or anti-CD10 antibodies were used simultaneously. In contrast to CD34-expressing cells, however, no consistent double-labeled subpopulation of lymphocytic cells was detected. Furthermore, coexpression of CD38 (73% ± 14%, n=4) or CD33 (29% ± 12%, n=5) on a majority of c-kit-positive cells showed their lineage commitment to erythropoiesis and granulocytopoiesis.

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