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Characterization of murine CD34, a marker for hematopoietic progenitor and
stem cells
DS Krause, T Ito, MJ Fackler, OM Smith, MI Collector, SJ Sharkis and WS May
Experimental Hematopoiesis Program, Johns Hopkins Oncology Center,
Baltimore, MD.
CD34 is expressed on human hematopoietic stem and progenitor cells, and its
clinical usefulness for the purification of stem cells has been well
established. However, a similar pattern of expression for murine CD34
(mCD34) has not yet been determined. Two polyclonal anti-mCD34 antibodies
that specifically recognize both endogenous and recombinant murine CD34
were developed to characterize the mCD34 protein and to determine its
pattern of expression on murine cell lines and hematopoietic progenitor
cells. Fluorescence-activated cell sorter analysis showed that mCD34 is
expressed on NIH/3T3 embryonic fibroblasts, PA6 stromal cells, embryonic
stem cells, M1 leukemia cells, and a subpopulation of normal bone marrow
cells. Murine CD34 was found to be a glycoprotein expressed on the cell
surface as either a full-length (approximately 100 kD) or truncated
(approximately 90 kD) protein in NIH/3T3 and PA6 cells. Recombinant
full-length CD34, when expressed in the CHO-K1 cell line, had a molecular
weight of approximately 105 kD. Full-length CD34 expressed on M1 leukemia
cells, had a higher apparent molecular weight (110 kD). These results
suggest that there are glycosylation differences between CD34 expressed by
different cell types. The full-length form, but not the truncated form, is
a phosphoprotein that is hyperphosphorylated in response to 12-0-
Tetradecanoyl phorbol 13-acetate treatment, suggesting potential functional
differences between the two forms. Selection of the 3% highest-expressing
CD34+ bone marrow cells enriched for the hematopoietic precursors that form
colony-forming unit-spleen (CFU-S), CFU-granulocyte-macrophage, and
burst-forming unit-erythroid. Transplantation of lethally irradiated mice
with these cells demonstrated both short- and long-term repopulating
ability, indicating that this population contains both functional
hematopoietic progenitors and the putative stem cell. These antibodies
should be useful to select for murine hematopoietic stem cells.
Volume 84,
Issue 3,
pp. 691-701,
08/01/1994
Copyright © 1994 by The American Society of Hematology
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