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Individual and combined effects of mesenchymal stromal cells and recombinant stimulatory cytokines on the in vitro growth of primitive hematopoietic cells from human umbilical cord blood

2009; Elsevier BV; Volume: 11; Issue: 7 Linguagem: Inglês

10.3109/14653240903180076

1477-2566

Patricia Flores-Guzmán, Eugenia Flores‐Figueroa, Juan José Montesinos, Guadalupe Martı́nez-Jaramillo, Verónica Fernández-Sánchez, Ignacio Valencia‐Plata, Guadalupe Alarcón-Santos, Héctor Mayani,

Neonatal Respiratory Health Research

Background aims We have previously characterized the in vitro growth of two cord blood-derived hematopoietic cell populations in liquid cultures supplemented with recombinant cytokines. In the present study, we assessed the effects of bone marrow-derived mesenchymal stromal cells (MSC) on the growth of such cells. Methods CD34 + CD38 + Lin − and CD34 + CD38 − Lin − cells were obtained by negative selection, and cultured in the presence of marrow-derived MSC and/or early- and late-acting cytokines. Hematopoietic cell growth was assessed throughout a 30-day culture period. Results In the presence of MSC alone, both populations showed significant proliferation. Direct contact between MSC and CD34 + cells was fundamental for optimal growth, especially for CD34 + CD38 − Lin − cells. In the presence of early-acting cytokines alone, cell growth was significantly higher than in cultures established with MSC but no cytokines. In cultures containing both MSC and early-acting cytokines, a further stimulation was observed only for CD34 + CD38 − Lin − cells. The cytokine cocktail containing both early- and late-acting cytokines was significantly more potent at inducing hematopoietic cell growth than the early-acting cytokine cocktail. When cultures were supplemented with early- and late-acting cytokines, MSC had no further effect on the growth of hematopoietic cells. Conclusions MSC seem to play a key role, particularly on more primitive (CD34 + CD38 − Lin − ) cells, only in the absence of cytokines or the presence of early-acting cytokines. When both early- and late-acting cytokines are present in culture, MSC seem to be unnecessary for optimal development of CFC and CD34 + cells.