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Hydrocortisone differentially affects the ability of murine stromal cells
and human marrow-derived adherent cells to promote the differentiation of
CD34++/CD38- long-term culture-initiating cells
L Croisille, I Auffray, A Katz, B Izac, W Vainchenker and L Coulombel
INSERM U 362, Institut Gustave Roussy, Villejuif, France.
Very primitive human hematopoietic progenitor cells are identified
indirectly by their ability to give rise to clonogenic progenitors in the
presence of either human or murine stromal cells. These long-term
culture-initiating cell (LTC-IC) assays are usually performed in the
presence of hydrocortisone based on the initial observation that
hydrocortisone was required for prolonged hematopoiesis in standard
long-term bone marrow cultures. In this report, we investigated the role of
hydrocortisone in LTC-IC assays initiated with CD34++/CD38- cells seeded
onto either human bone marrow LTC-derived adherent cells or a murine
marrow-derived stromal cell line, MS-5. It was found that weekly addition
of hydrocortisone to the cultures reduced the frequency of LTC-IC (from 1/5
to 1/20) calculated from limiting dilution experiments and also reduced
fivefold to 10-fold the number of their progeny clonogenic cells detected
after 4 to 5 weeks. In contrast, the frequency and differentiative
potential of CD34++/CD38- grown in the presence of human marrow feeders was
unaltered by the addition of glucocorticoids. Data are consistent with the
hypothesis that hydrocortisone inhibited LTC-IC differentiation by
downregulating the expression of a synergistic factor produced by MS-5
cells. (1) In the absence of hydrocortisone, the number of clonogenic
progenitors generated by LTC-IC was much higher in cultures seeded on MS-5
than in cultures seeded on human marrow adherent cells, which was also true
when cytokines were added to the cocultures. However, based on the
phenotype of the colonies, progenitors produced in MS-5 cocultures were
more mature than those generated on human marrow adherent cells. (2)
Hydrocortisone counteracted the stimulatory effect of recombinant human
cytokines (interleukin-3, interleukin-6, and steel factor) in assays
performed on MS-5 but not on human marrow feeders. (3) Hydrocortisone led
to a 50% decrease in the numbers of colony-forming units-
granulocyte-macrophage found in methycellulose colony assays of
CD34++/CD38- cells performed in the presence of MS-5 cells. Taken together,
our results indicate that hydrocortisone acts differently on a murine
stromal cell line and on marrow-derived human stromal cells and may
suppress the expression by MS-5 cells of an activity selectively promoting
amplification of clonogenic cells derived from primitive LTC-IC.
Volume 84,
Issue 12,
pp. 4116-4124,
12/15/1994
Copyright © 1994 by The American Society of Hematology
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