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Blood, 15 January 2001, Vol. 97, No. 2, pp. 435-441
HEMATOPOIESIS
Stromal cells retard the differentiation of
CD34+CD38low/neg human primitive progenitors
exposed to cytokines independent of their mitotic history
Annelise Bennaceur-Griscelli,
Corinne Pondarré,
Valérie Schiavon,
William Vainchenker, and
Laure Coulombel
From INSERM U 362, Institut Gustave Roussy, Villejuif,
and INSERM U 474, Hôpital Port Royal, Paris, France.
Stem cell proliferation induced by potent cytokines usually leads
to a loss of primitive potential through differentiation. In this
study, the ability of cytokines and murine MS5 stromal cells to
independently regulate the proliferation and long-term culture-initiating cell (LTC-IC) activity of primitive
CD34+CD38low/neg human bone marrow cells was
evaluated. To compare populations with identical proliferation
histories, cells were labeled with carboxy fluorescein diacetate
succinimidyl ester, and LTC-IC activity was assessed 4 days later in
cells that had accomplished the same number of divisions with or
without MS5 cells. MS5 cells counteracted dramatically the loss of
LTC-IC activity observed in the presence of cytokines alone. Thus, in
the presence of MS5 cells, means of 1233 (n = 5) and 355 (n = 9)
LTC-IC-derived colony-forming cells (CFCs) were generated by 1000 cells that performed 3 and 4 divisions respectively, whereas 311 (n = 5) and 64 (n = 5) CFCs were generated by 1000 cells cultured
without MS5 cells. Interestingly, MS5 cells had no detectable effect on
the LTC-IC activity of cells that divided only twice in 4 days 1606
CFCs (n = 6) and 1993 (n = 6) CFCs, respectively, without and with
MS5 cellsand a 48 additional hours of coculture were necessary to
unmask changes in the LTC-IC activity mediated by stromal cells. These
results indicate that cytokines and stroma-derived signals can
regulate independently the proliferation and differentiation of
primitive cells and that these stroma-derived extracellular factors act
directly on their target cells.
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