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Cell cycle and functional differences between CD34+/CD38hi and CD34+/38lo
human marrow cells after in vitro cytokine exposure
JA Reems and B Torok-Storb
Clinical Research Division, Fred Hutchinson Cancer Research Center,
Seattle, WA 98104.
The proliferation kinetics and clonogenic activity of CD34+/38hi (CD38hi)
and CD34+/38lo (CD38lo) human marrow cells were measured before and after
culturing the cells in vitro over a 6-day period in serum-deprived medium
containing recombinant growth factors (interleukin-1 [IL-1], IL-3, IL-6,
granulocyte colony-stimulating factor [G-CSF], granulocyte-macrophage
[GM]-CSF, kit ligand, and erythropoietin). Before in vitro culture, 3% +/-
3% of the CD38lo and 13% +/- 2% of the CD38hi cells were in the S-phase of
the cell cycle. The clonogenic activity of CD38hi cells was twofold greater
than that of the CD38lo cells, as measured by colony-forming units (CFU) in
short- term assays. However, CD38hi cells contained fewer pre-CFU than did
the CD38lo cells, generating only 3 +/- 2 colonies per 1,000 cells after 4
weeks of culture on competent stromal layers, compared with 107 +/- 46
colonies per 1,000 cells from the CD38lo population. CD38hi and CD38lo
cells exhibited distinctly different responses when cultured in serum-
deprived medium supplemented with recombinant growth factors. After
culturing cells for 24 hours, CD38lo cells essentially remained a
noncycling population with only 5.1% +/- 3.0% of the cells cycling, whereas
44.2% +/- 6.9% of the CD38hi cells were in DNA synthesis. Gradually CD38lo
cells were recruited into cycle, such that by 72 hours, approximately 28%
of the CD38lo cells were in S-phase. However, during 6 days of culture, the
percentage of cycling CD38lo cells never exceeded the proliferative
response observed for CD38hi cells. Phenotype analysis conducted at day 6
indicated that 86% of the CD38hi population were no longer phenotypically
CD34+/38hi, while 60% of CD38lo cells maintained a CD34+/38lo phenotype.
Long-term cultures initiated with 6-day in vitro-expanded CD38lo cells
showed approximately a twofold decrease in clonogenic activity attributable
to a loss of erythroid precursors and a decrease in GM colonies. Thus, a
proportion of CD38lo cells capable of generating CFU was maintained even
after exposure to growth factors.
Volume 85,
Issue 6,
pp. 1480-1487,
03/15/1995
Copyright © 1995 by The American Society of Hematology
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