Effect of allogeneic adipose derived mesenchymal stem cells on regulatory T cells in peripheral blood of patients with premature ovarian failure

Objective To investigate the effect of allogeneic adipose derived mesenchymal stem cells (hADSCs) on regulatory T cells (Tregs) in peripheral blood of patients with premature ovarian failure (POF). Methods hADSCs were isolated from adipose tissue using 0.1% type Ⅰ collagenase digestion. Adipogenic, osteogenic differentiation and surface molecular characterization were also performed. Peripheral blood mononuclear cells (PBMCs) of POF patients were isolated by density gradient centrifugation. PBMCs were co-cultured with 1×104, 2×104 and 1×105 hADSCs for 72 hours under the stimulation of phytohemagglutinin (PHA). The proliferation rate of lymphocytes was measured by CCK-8 method, the proportion of CD4+CD25+Foxp3+ Treg cells was measured by flow cytometry. Real-time fluorescent quantitative PCR was used to detect the mRNA level of Foxp3+. Results hADSCs were positive for CD90, CD105 and negative for CD34, CD45, and has adipogenic and osteogenic differentiation ability. The results of CCK-8 showed that hADSCs could significantly inhibit the proliferation of lymphocytes compared with that in control group (P<0.001). Flow cytometry showed that hADSCs could promote the proliferation of CD4+CD25+Foxp3+ Treg cells (P<0.05). The results of qPCR showed that Foxp3+ mRNA expression was obviously up-regulated in all experimental groups compared with that in control group (P<0.001). Conclusion hADSCs can play an immune regulatory role and promote the proliferation of Tregs in peripheral blood of patients with premature ovarian failure. DOI: 10.11855/j.issn.0577-7402.2018.04.05