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N Sato, K Sawada, M Kannonji, T Tarumi, N Sakai, M Ieko, S Sakurama, S Nakagawa, T Yasukouchi and SB Krantz
Second Department of Internal Medicine, Hokkaido University School of
Medicine, Sapporo.
To facilitate the investigation of the direct interaction between
hematopoietic progenitors and colony-stimulating factors, we have developed
a method to purify human marrow progenitor cells. Using density
centrifugation, negative panning with concanavalin A coated plates,
positive selection of CD34-positive cells with immunomagnetic microspheres,
overnight adherence to a plastic dish, negative selection with a panel of
monoclonal antibodies, and density centrifugation, human marrow progenitor
cells were purified from 1.5% to 53.2%, a 42- fold purification, with a
4.8% yield. The purified cells consisted of 38% erythroid, 9% colony
forming unit-granulocyte (CFU-G), 29% CFU- macrophage (CFU-M), 12%
CFU-eosinophil/basophil (CFU-Eo/Ba), and 4% CFU- mix. The purified cells
cultured in serum-free fibrin clots with recombinant human macrophage
colony-stimulating factor (rM-CSF) for 14 days developed a pure population
of CFU-M colonies. An appearance of CFU-M colonies was present after the
addition of 1 U/mL of rM-CSF and the maximum stimulation was found at 100
U/mL. When the purified cells were cultured in serum-free medium with
rM-CSF in a limiting dilution assay and the percentage of nonresponder
wells for CFU-M colonies was plotted against cell concentration, serum-free
cultures yielded a straight line through the origin, indicating that CFU-M
development did not depend on accessory cells and that rM-CSF acted
directly on the CFU- M.
This article has been cited by other articles:
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| Copyright © 1991 by American Society of Hematology Online ISSN: 1528-0020 | |||||||||
