Monoclonal antibodies against human hemopoietic cells and the separation of progenitor cells from bone marrow.
Monoclonal antibodies against myeloid cell surface antigens were selected according to their reactivity against normal human bone marrow cells. Each of the antibodies detected a different population of maturing bone marrow cells, but almost none of the progenitor cells assayed (seven-day GM-CFC, 14-day GM-CFC, CFU-E, BFU-E). Myeloid cells, erythroid cells, and the majority of lymphocytes and monocytes were simultaneously depleted with the mixture of these three antibodies using fluorescence-activated cell sorting (FACS) or panning for cell separation; 14-day GM-CFC and BFU-E were enriched six- to 16-fold when negative cells were sorted. After a negative selection step using panning, BFU-E and 14-day GM-CFC were enriched four- to seven-fold and five- to 13-fold, respectively. Negative cells obtained after panning or cell sorting were also enriched in blast cells (24.4% and 27.2%, respectively) and depleted in maturing bone marrow cells, with the exception of variable numbers of lymphocytes, monocytes, and plasma cells. FACS of negative cells using both forward light-scatter and perpendicular light-scatter parameters resulted in a cell population that contained a majority of undifferentiated blasts and 5.5%-9.6% BFU-E and 14-day GM-CFC.
D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001854:Bone Marrow Cells; D002469:Cell Separation; D003114:Colony-Forming Units Assay; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000276:immunology*; D006801:Humans
D000911:Antibodies, Monoclonal