Objective: To study the immunomodulatory functions of human unbilical cord mesenchymal stem cells on bone marrow T-lymphocyte of immune mediated aplastic anemia.Methods: MSCs were isolated from normal human unbilical cord,cultured in vitro. T lymphocytes were isolated from bone marrow of immune mediated aplastic anemia by density gradient centrifugation method. The MSCs and T lymphoeytes were co-cultured. The proliferation of lymphocytes was measured by MTT method, the phenotypes of the T cells were analysed by flow cytometry before and after coculture with MSC and the expression of the two marrow hematopoietic factors of aplastic anemia were evaluated by ELISA.Results: (1) In intro,HUC-MSCs were generated successfully. The second generation of MSCs had tyical morphology of fiberblast,and were high expression of CD166,CD29,CD54,low expression of CD13,CD34,CD45 . MSCs could be induced to adipocyte. (2) HUC-MSCs showed inhibitory effect on LCs of aplastic anemia proliferation when HUCMSCs and LCs mixed at the ratio of 1:1,0.5:1,0.1:1, and their inhibitory rate were 56.2±12.1%,43.7±10.4%,28.6±8.9%,3.41±7.6%; HUCMSCs can adjust the proportion of CD4+ and CD8+ T cell subset; and HUCMSCs also can inhibit the secretion of IL-2 and IFN-γby T cells of aplastic anemia.Conclusion:Human unbilical cord mesenchymal stem cells can mediate a immunregulatory effect of T lymphocyte of immune mediated aplastic anemia in vitro and the inhibitory effects are dependent on the amount of HUC-MSCs.
|