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Identification and comparison of CD34-positive cells and their
subpopulations from normal peripheral blood and bone marrow using
multicolor flow cytometry
JG Bender, KL Unverzagt, DE Walker, W Lee, DE Van Epps, DH Smith, CC Stewart and LB To
Applied Sciences, Baxter Healthcare Corporation, Round Lake, IL.
Four-color flow cytometry was used with a cocktail of antibodies to
identify and isolate CD34+ hematopoietic progenitors from normal human
peripheral blood (PB) and bone marrow (BM). Mature cells that did not
contain colony forming cells were resolved from immature cells using
antibodies for T lymphocytes (CD3), B lymphocytes (CD20), monocytes (CD14),
and granulocytes (CD11b). Immature cells were subdivided based on the
expression of antigens found on hematopoietic progenitors (CD34, HLA-DR,
CD33, CD19, CD45, CD71, CD10, and CD7). CD34+ cells were present in the
circulation in about one-tenth the concentration of BM (0.2% v 1.8%) and
had a different spectrum of antigen expression. A higher proportion of
PB-CD34+ cells expressed the CD33 myeloid antigen (84% v 43%) and expressed
higher levels of the pan leukocyte antigen CD45 than BM-CD34+ cells. Only a
small fraction of PB-CD34+ cells expressed CD71 (transferrin receptors)
(17%) while 94% of BM-CD34+ expressed CD71+. The proportion of PB-CD34+
cells expressing the B-cell antigens CD19 (10%) and CD10 (3%) was not
significantly different from BM-CD34+ cells (14% and 17%, respectively).
Few CD34+ cells in BM (2.7%) or PB (7%) expressed the T-cell antigen CD7.
CD34+ cells were found to be predominantly HLA-DR+, with a wide range of
intensity. These studies show that CD34+ cells and their subsets can be
identified in normal PB and that the relative frequency of these cells and
their subpopulations differs in PB versus BM.
Volume 77,
Issue 12,
pp. 2591-2596,
06/15/1991
Copyright © 1991 by The American Society of Hematology
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