Characterization of hemopoietic cell populations from human cord blood expressing c-kit
| Autor: | G, Reisbach, I, Bartke, B, Kempkes, G, Kostka, J, Ellwart, A, Birner, K, Thalmeier, R, Mailhammer, G W, Bornkamm, A, Ullrich |
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| Rok vydání: | 1993 |
| Předmět: |
B-Lymphocytes
Membrane Glycoproteins Macrophages T-Lymphocytes Sialic Acid Binding Ig-like Lectin 3 Antibodies Monoclonal Antigens Differentiation Myelomonocytic Fluorescent Antibody Technique Antigens CD34 Bone Marrow Cells Cell Separation Fetal Blood Flow Cytometry Hematopoietic Stem Cells ADP-ribosyl Cyclase 1 Antigens Differentiation Proto-Oncogene Proteins c-kit Antigens CD Proto-Oncogene Proteins Humans ADP-ribosyl Cyclase Cells Cultured Granulocytes |
| Zdroj: | Experimental hematology. 21(1) |
| ISSN: | 0301-472X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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