Human interleukin for DA cells (HILDA) does not affect the proliferation and differentiation of hematopoietic progenitor cells in human long-term bone marrow cultures

Abstract

Human Interleukin for DA cells (HILDA), a cytokine also known as leukemia inhibitory factor (LIF), induces proliferation without concurrent differentiation of murine embryonic stem cells. Therefore, we investigated the effects of recombinant HILDA/LIF on the proliferation and differentiation of human hematopoietic progenitor cells (HPC) grown in long-term bone marrow cultures (LTBMC). Pre-established stromal cell layers were reinoculated with autologous cryopreserved mononuclear phagocyte- and T-lymphocyte-depleted bone marrow cells in the presence or absence of HILDA/LIF (200 U/ml). At weekly intervals cultures were sacrificed, and the cells in the adherent and the nonadherent cell fractions were counted. The numbers of HPC were determined by culturing these cells in semisolid medium stimulated with phytohemagglutinin-stimulated leukocyte-conditioned medium (PHA-LCM), and LTBMC supernatants were assayed in semisolid cultures for the presence of colony-stimulating activity (CSA). The total number of cells, their differential counts, the number of HPC, and the concentrations of CSA in culture supernatants were similar for long-term cultures containing HILDA/LIF and for controls. These data suggest that HILDA/LIF may not play a role in the proliferation and differentiation of normal human (early) HPC in LTBMC. Moreover, HILDA/LIF did not stimulate the proliferation of relatively mature progenitor cells in semisolid cultures, not did it influence the colony formation induced by other colony-stimulating factors (CSF). Finally, using a [3H]thymidine suicide test we could not find an effect of HILDA/LIF on the cell-cycle status of HPC.