Enrichment of pluripotent hemopoietic progenitor cells from human bone
marrow
MP Bodger, IM Hann, RF Maclean and ME Beard
Pluripotent hemopoietic progenitor cells (CFU-GEMM, cells forming mixed
hemopoietic colonies in methylcellulose) from human bone marrow were
enriched 90-fold by positive selection on the fluorescence-activated cell
sorter using monoclonal antibody RFB-1. Bone marrow cells were separated by
cell size, using log 90 degrees light scatter, and the cell fraction
containing CFU-GEMM was further separated by relative fluorescence
intensity for the RFB-1 antigen. Further enrichment, up to 150-fold, was
achieved by depleting bone marrow of T cells and mature myeloid cells prior
to RFB-1 selection. These procedures yield a cell fraction containing 51%
blast cells, 2% promyelocytes, and 47% undifferentiated (lymphocyte-like)
mononuclear cells, although only 1% of the cells formed a mixed colony.
CFU-GEMM are strongly positive for the RFB-1 antigen, whereas
morphologically identifiable erythroblasts, myeloblasts, and promyelocytes
are weakly RFB-1+. This suggests that the relative concentration of the
RFB-1 antigen on bone marrow cells is inversely related to their maturity.
The greatly increased recovery of CFU-GEMM after the separation of bone
marrow by log 90 degrees light scatter and the removal of T cells and
mature myeloid cells suggested that accessory cells that normally regulate
the cloning efficiency of CFU-GEMM were removed.
Volume 64,
Issue 4,
pp. 774-779,
10/01/1984
Copyright © 1984 by The American Society of Hematology
