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目的观察脐带间充质干细胞(UC-MSC)对体外外周血单核细胞的影响。方法分离人UC-MSC和外周血单核细胞,流式分析技术鉴定单核细胞CD45和CD3的表达率;UC-MSC和单核细胞按不同比例共培养72 h,采用免疫荧光染色和ELISA方法检测γ干扰素(IFN-γ)在细胞内和培养上清的表达。结果采用含10%血清的DMEM/F12可以从新鲜脐带分离到UC-MSC,密度梯度离心法可以从外周血中分离到单核细胞,CD45阳性率为89.4%,CD3阳性率为80.7%。可在CD3/CD28刺激培养的单核细胞检测到IFN-γ的表达,1∶10和1∶20共培养上清IFN-γ的含量显著低于仅有单核细胞的对照组(P<0.01)。结论UC-MSC的存在可以影响IFN-γ在单核细胞的表达,这种影响与两种细胞的比例有关。
Objective To observe the effect of umbilical cord mesenchymal stem cells (UC-MSCs) on peripheral blood mononuclear cells in vitro. Methods UC-MSCs and peripheral blood mononuclear cells were isolated by flow cytometry. The expression rates of CD45 and CD3 on mononuclear cells were determined by flow cytometry. UC-MSCs and mononuclear cells were co-cultured for 72 h at different ratios. Immunofluorescence staining and ELISA The expression of interferon-γ (IFN-γ) in cells and culture supernatants was detected. Results UC-MSCs could be isolated from fresh umbilical cord by DMEM / F12 containing 10% serum. Mononuclear cells could be isolated from peripheral blood by density gradient centrifugation. The positive rate of CD45 was 89.4% and the positive rate of CD3 was 80.7%. The expression of IFN-γcan be detected in CD3 / CD28-stimulated monocytes. The content of IFN-γ in 1:10 and 1:20 co-culture supernatant was significantly lower than that in monocyte-only control group (P <0.01 ). Conclusions The presence of UC-MSCs can affect the expression of IFN-γ in monocytes. This effect is related to the ratio of the two types of cells.