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目的研究人脐带间充质干细胞(huMSCs)向胰岛素分泌细胞(IPCs)分化过程中的免疫学特性,为huMSCs作为胰岛细胞移植治疗1型糖尿病的细胞来源提供依据。方法以流式细胞术检测huMSCs及huMSCs源性IPCs的免疫表型;RT-PCR法检测huMSCs及huMSCs源性IPCs的HLA-A、HLA-DR基因的表达;CCK8法测定细胞增殖率,观察huMSCs及huMSCs源性IPCs对外周血单个核细胞增殖的影响。结果 huMSCs在体外培养条件诱导下,具有向胰岛素分泌细胞分化的潜能;huMSCs及huMSCs源性IPCs均不表达CD80、CD86、CD40、CD40L、HLA-DR,均表达HLA-A;huMSCs能够抑制PHA刺激的外周血单个核细胞的增殖,但huMSCs源性IPCs未见该作用。结论 huMSCs及huMSCs源性IPCs均具有低免疫原性,可作为胰岛细胞移植的细胞来源,huMSCs对免疫反应具有负调节作用,但huMSCs源性IPCs不具有该作用。
Objective To study the immunological characteristics of human umbilical cord mesenchymal stem cells (hMSCs) differentiating into insulin-secreting cells (IPCs) and to provide basis for hMSCs as the source of cells for type 1 diabetes mellitus after islet cell transplantation. Methods The immunophenotypes of huMSCs and huMSCs derived IPCs were detected by flow cytometry. The expression of HLA-A and HLA-DR genes in huMSCs and huMSCs derived IPCs were detected by RT-PCR. The proliferation of huMSCs was detected by CCK8 assay. And huMSCs-derived IPCs on the proliferation of peripheral blood mononuclear cells. Results huMSCs had the potential to differentiate into insulin-secreting cells under the condition of in vitro culture. The huMSCs and huMSCs derived IPCs did not express CD80, CD86, CD40, CD40L and HLA-DR, all expressed HLA-A. Peripheral blood mononuclear cells proliferated, but this effect was not seen with huMSCs derived IPCs. Conclusions Both huMSCs and huMSCs derived IPCs have low immunogenicity and can be used as the source of cells for islet cell transplantation. The huMSCs have a negative regulatory effect on immune responses, but huMSCs derived IPCs do not have this effect.