The Different Immunomodulatory Of The Mesenchymal Stem Cells From Itp Patients Bone Marrow And Other Sources

AIM In this research,we will co-culture T lymphocytes from normal humans with MSCs which were isolated from Immune thrombocytopenia patients’s bone marrow, Umbilical cord and healthy people’s bone marrow.3days later, we will monitor the cytokines IL-2, IFN-y, IL-4and IL-10to illustrate the diffence of immunodulatory of three kinds of MSCs.Methods:Human bone marrow-derived MSCs were isolated by Ficoll-Pague density gradient centrifugation from Immune thrombocytopenia patients (ITP)(n=16) and healthy people (n=8) and cultured for5-6passages, and then the cells were collected and used for the study. Umbilical cord mesenchymal stem cells (UC-MSCs) were isolated from10healthy fetal umbilical cord and the passage5-6cells were used. The mononuclear cells were isolated from healthy adult’s peripheral blood by Ficoll-Hypaque gradient centrifugation. The three kinds of MSCs were inactivated by mitomycin C and then were co-cultured with PHA-activated T cells with the ratio of1:40,3:40,and 9:40respectively.3days later, the culture supernatant was collected and the cytokines including INF-γ, IL-2, IL-4and IL-10were measured by ELISA.bResults The characteristics of ITP patients’s BMMSCs were similar with that of healthy donor MSCs and UC-MSCs, the cells exhibited spindle-like morphology. MSCs from acute ITP patients’bone marrow showed a similar growth rate with that of the healthy donor MSCs, but MSCs from chronic ITP patients showed a slower growth rate. MSCs of different origin showed similar phonotype, which were positive for CD29, CD13, CD44, CD90and CD105, while negative for CD34, CD45and HLA-DR. They have the same capacity of both adipocytes and osteoblasts differentiation. All the MSCs inhibited the proliferation of PHA-activated T cells in a dose-dependent way. When the mononuclear cells were co-cultured with inactivated MSCs of different origin, more MSCs resulted in the increase of IL-10and decrease of IL-2, INF-γ and IL-4in the supernatent. While there was no difference in the secretion of cytokines between MSCs of different origin.Conclusion:(1)BMMSCs from ITP patients exhibited the same characterics and phonotye with healthy donor MSCs and UC-MSCs, but showed a slower growth rate.(2)When the mononuclear cells were co-cultured with inactivated MSCs, the secretion of IL-2, IFN-γ and IL-4are proportional to the MSCs number, IL-10secretion is inversely proportional to the MSCs number.(3)There was no significant difference in the secretion of IL-2, IFN-γ, IL-4and IL-10between MSCs of different origin.