Effect of allogeneic adipose derived mesenchymal stem cells on regulatory T cells in peripheral blood of patients with premature ovarian failure / 解放军医学杂志

ABSTRACT

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.