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中国应用生理学杂志 ›› 2018, Vol. 34 ›› Issue (5): 396-400.doi: 10.12047/j.cjap.5621.2018.090

• 研究论文 • 上一篇    下一篇

HLA-G阳性的胎盘间充质干细胞体外诱导Treg的实验研究

崔桂玉1, 白剑2, 苗兰英2, 林大勇2, 刘鸿2, 李亚丽3,4, 刘喜成3,4   

  1. 1. 内蒙古民族大学医学院, 通辽 028000;
    2. 辽宁中医药大学, 沈阳 110847;
    3. 深圳市人民医院麻醉科, 广东 深圳 518020;
    4. 深圳市麻醉医学工程研究中心, 广东 深圳 518020
  • 收稿日期:2017-07-27 修回日期:2018-02-11 出版日期:2018-09-28 发布日期:2019-02-21
  • 通讯作者: 刘喜成,Tel:0755-25533018;E-mail:flameliu@sina.com E-mail:flameliu@sina.com
  • 基金资助:
    国家自然科学基金面上项目(81571555);辽宁省自然科学基金(2015020714)

Placenta-derived mesenchymal stem cells with HLA-G positive expression induce Treg in vitro

CUI Gui-yu1, BAI Jian2, MIAO Lan-ying2, LIN Da-yong2, LIU Hong2, LI Ya-li3,4, LIU Xi-cheng3,4   

  1. 1. Medical School of Inner Mongolia University for Nationalities, Tongliao 028000;
    2. Liao Ning University of Traditional Chinese Medicine, Shenyang 110847;
    3. Department of Anesthesiology, Shenzhen People's Hospital, Shenzhen 518020;
    4. Shenzhen Anesthesiology Engineering Center, Shenzhen 518020, China
  • Received:2017-07-27 Revised:2018-02-11 Online:2018-09-28 Published:2019-02-21
  • Supported by:
    国家自然科学基金面上项目(81571555);辽宁省自然科学基金(2015020714)

摘要: 目的:研究HLA-G阳性的胎盘间充质干细在体外诱导Treg的产生。方法:从新生儿胎盘中分离胎盘间充质干细胞的,采用脂质体转染的方式将PEGFP-N1-HLA-G质粒转染到胎盘间充质干细胞中,将细胞分为空白对照组、PEGFP-N1组和PEGFP-N1-HLA-G组,每组设置5个复孔,并通过蛋白质免疫印迹检测HLA-G的表达,将鉴定后的细胞与健康人外周血中CD4+的T淋巴细胞混合培养24 h和48 h,并检测CD4+CD25+Foxp3+Treg占T淋巴细胞的比例。结果:PEGFP-N1-HLA-G转染后胎盘间充质干细胞可以表达HLA-G蛋白,与空白对照组和PEGFP-N1组相比有显著性差异(P<0.01);HLA-G阳性的胎盘间充质干细胞在与CD4+的T淋巴细胞混合淋巴细胞培养24 h后,Treg细胞占全部T淋巴细胞的比例为(16.41±0.94)%,在培养48 h后,Treg细胞的占全部T淋巴细胞的比例为(16.46±0.59)%,与空白对照组和PEGFP-N1组相比有显著性差异(P<0.01)。结论:HLA-G基因修饰后胎盘间充质干细胞能够有效的在体外诱导CD4+ CD25+ FoxP3+Treg产生。

关键词: HLA-G, 胎盘间充质干细胞, T淋巴细胞, 调节性T淋巴细胞

Abstract: Objective: To study placenta-derived mesenchymal stem cells with HLA-G (Human Leukocyte Antigen, HLA-G) positive expression induce Treg (regulatory T cell, Treg) in vitro. Methods: placenta-derived mesenchymal stem cells were separated from neonatal placenta; PEGFP - N1 -HLA-G plasmid was transfected in placenta-derived mesenchymal stem cells by liposome transfection.The cells were divided into 3 groups including control group, PEGFP-N1 group and PEGFP-N1-HLA-G group, 5 complex walls in each group. Expression of HLA-G protein was detected by Western Blotting; after identification of cells, healthy human peripheral blood CD4+ T lymphocytes were cultured with placenta-derived mesenchymal stem cells with HLA-G positive expression, and the ratio of CD4+CD25+Foxp3+Treg in T lymphocytes was accounted. Results: After transfection of PEGFP-N1-HLA-G, the placenta-derived mesenchymal stem cells can express HLA-G protein significantly, compared with the control group and PEGFP - N1 group (P<0.01). After HLA-G positive placenta-derived mesenchymal stem cells and CD4 + T lymphocytes were cultured for 24 h, the ratio of CD4+CD25+Foxp3+Treg in T lymphocytes was (16.41±0.94)%. After HLA - G positive placenta-derived mesenchymal stem cells and CD4+ T lymphocytes were cultured for 48 h, the ratio of CD4+CD25+Foxp3+Treg in T lymphocytes was (16.46±0.59)% significantly, compared with the control group and PEGFP - N1 group (P<0.01). Conclusion: Placenta-derived mesenchymal stem cells modified by HLA-G gene can effectively induce CD4+CD25+Foxp3+Treg in vitro.

Key words: human leucocyte antigen-G, placenta-derived mesenchymal stem cell, T lymphocyte, regulatory T lymphocyte

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