Monoclonal antibodies against human hemopoietic cells and the separation of progenitor cells from bone marrow
Mouchiroud, G.; Berthier, R.; Newton, I.A.; Chapel, A.; Hollard, D.
Experimental Hematology 13(6): 566-573
1985
ISSN/ISBN: 0301-472X PMID: 3996493 Document Number: 256740
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 (7-day GM-CFC, 14-day GM-CFC ). 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 6- to 16-fold when negative cells were sorted. After a negative selection step using panning, BFU-E and 14-day GM-CFC were enriched 4- to 7-fold and 5- 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.