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脐带间充质干细胞对体外外周血单核细胞的影响

ISSN号：1004-0188
期刊名称：西南国防医药
时间：2015.8.20
页码：820-822
分类：R318.1[医药卫生—生物医学工程;医药卫生—基础医学]
作者机构：[1]成都军区昆明总医院细胞生物治疗中心、干细胞与免疫细胞生物医药技术国家地方联合工程实验室、云南省细胞治疗技术转化医学重点实验室、云南省干细胞工程实验室,昆明650032
相关基金：云南省基金项目（2011HB050,2013DA004,2013CA005）;成都军区基金项目（2012WJ0003）;国家自然科学基金项目（31172170）
相关项目：Ⅱ型糖尿病的猕猴模型构建及重编程干细胞的效应机制

关键词：脐带, 间充质干细胞, 单核细胞, 共培养, Γ干扰素, 影响, umbilical cord, mesenchymal stem cell, mononuclear cell, co-culture, interferon-γ, effect

中文摘要：

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。

英文摘要：

Objective To observe the effects of umbilical cord mesenchymal stem cells （UC-MSC） on mononuclear cells of peripheral blood in vitro. Methods Human UC-MSC and mononuclear cells of peripheral blood were isolated. Flow cytometry was used to identify the expression rate of mononuclear cell CD45 and CD3. UC-MSC and mononuclear ceils were co-cultured in different proportion for 72 h. Immunofluorescence staining and ELISA were used to detect the expression of interferon-γ （IFN-γ） within the ceils and the ceU culture supernatant, Results UC-MSC could be isolated from fresh umbihcal cord by DMEM/F12 containing 10% serum. Mononuclear cells could be isolated from the peripheral blood by density gradient centrifugation. The positive rate of CD45 and CD3 was 89.4% and 80.7%, respectively. The expression of IFN-γ was detected in CD3/CD28-stimulated- mononuclear cells. The IFN-γ content in the co-cultured supernatant in the proportion of 1：10 and 1：20 was significantly lower than that in the control group with only mononuclear cells （P 〈 0.01）. Conclusion The presence of UC-MSC can affect the expression of IFN-γ in mononuclear cells, and the effects are related to the proportion of UC-MSC and mononuclear cells.

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