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Rapid discrimination of early CD34+ myeloid progenitors using CD45-RA
analysis
G Fritsch, P Buchinger, D Printz, FM Fink, G Mann, C Peters, T Wagner, A Adler and H Gadner
Children's Cancer Research Institute, Vienna, Austria.
Mononuclear cells (MNC) isolated by density centrifugation of cord blood
and healthy bone marrow, and of peripheral blood (PB) from patients treated
with granulocyte-macrophage colony-stimulating factor (GM-CSF) or G-CSF
after chemotherapy, were double-stained with anti CD34 monoclonal antibody
(MoAb) (8G12) versus anti CD45, CD45-RB, CD45- RO, and CD45-RA,
respectively, and analyzed by flow cytometry. In all specimens, CD34+ MNC
co-expressed CD45 at a low level and the expression of CD45-RB was similar
or slightly higher. Most CD34+ MNC were negative for CD45-RO, a weak
coexpression was only seen in some bone marrow (BM) and blood samples. In
contrast, CD45-RA could subdivide the CD34+ population into fractions
negative, dim (+), and normal positive (++) for these subgroups, and
typical staining patterns were observed for the different sources of
hematopoietic cells: in BM, most CD34+ MNC were RA++. In PB, their majority
was RA++ after G-CSF but RA+ or RA- after GM-CSF. In cord blood, the
hematopoietic progenitors were mainly RA-/RO-. Semisolid culture of sorted
CD34+ MNC showed that clusters and dispersed (late) colony-forming unit-GM
(CFU- GM) originated from 34+/RA++ cells, while the 34+/RA- MNC formed
compact and multicentric, both white and red colonies derived from early
progenitors. Addition of 20 ng stem cell factor per milliliter of medium
containing 34+/RA- cord blood MNC led to a change of many burst- forming
unit-erythrocyte (BFU-E) to CFU-mix which was not, at least to this extent,
seen in blood and BM. We conclude that early myeloid CD34+ cells are
45+/RA-. Because this population excludes 34+/19+ B cells and 33+ myeloid
cells, both of which are RA++, two-color flow cytometric analysis using
CD34 and CD45-RA facilitates the characterization and quantification of
early myeloid progenitor cells.
Volume 81,
Issue 9,
pp. 2301-2309,
05/01/1993
Copyright © 1993 by The American Society of Hematology
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