胎盘间充质干细胞培养上清对CD4^+T细胞和Treg细胞免疫功能的调节  

CD4^+ T and Treg immunomodulatory functions after culture with the supernatant of human placenta mesenchymal stem cells

作　　者：李伟伟 李晓丰[1] 侯萍 李剑平[1] Li Weiwei;Li Xiaofeng;Hou Ping;Li Jianping(Shenyang Blood Center(Liaoning Provincial Blood Center), Shenyang 110044,Liaoning Province,China)

机构地区：[1]沈阳中心血站(辽宁省血液中心),辽宁省沈阳市110044

出　　处：《中国组织工程研究》2019年第1期85-89,共5页Chinese Journal of Tissue Engineering Research

基　　金：2017年辽宁省博士科研启动基金指导计划项目(20170520339);项目负责人:李伟伟~~

摘　　要：背景:Treg细胞是一种抑制性CD4^+T细胞,人胎盘间充质干细胞具有T细胞免疫调节功能。目的:探讨人胎盘间充质干细胞上清液对CD4^+T细胞和Treg细胞比例以及免疫调节功能的影响。方法:分离培养人胎盘间充质干细胞,建立其稳定传代细胞系。收集第4代人胎盘间充质干细胞及细胞上清培养液,流式细胞术检测细胞表面标志物CD90、CD73、CD45、HLA-DR的表达。采用人胎盘间充质干细胞上清液与细胞因子刺激剂联合培养正常人外周血单个核细胞6 h,流式细胞术检测CD4^+T细胞和Treg细胞比例;ELISA方法检测细胞培养上清中转化生长因子β水平。结果与结论:(1)成功建立人胎盘间充质干细胞原代细胞系,且可稳定传代;(2)人胎盘间充质干细胞上清液能够显著降低CD4^+T细胞表达比例(P <0.05),增强Treg细胞的表达比例(P <0.05),并上调其主要细胞因子转化生长因子β水平(P <0.01);(3)结果表明,人胎盘间充质干细胞上清液可以通过诱导CD4^+T细胞向Treg细胞的转化而发挥免疫调节作用,有望成为一种免疫治疗的新选择。BACKGROUND:Treg cells belong to inhibitory CD4+T cells.Human placenta mesenchymal stem cells(hPAMSCs)have T cell immunomodulatory function.OBJECTIVE:To investigate the CD4+T and Treg cells immunomodulatory ability after culture with the supernatant of hPAMSCs.METHODS:hPAMSCs were isolated and cultured,and the stable cell lines were then established.Passage 4 hPAMSCs and its supernatant were collected,and the expression of cell surface markers CD90,CD73,CD45 and HLA-DR were detected by flow cytometry.Cell activation cocktail stimulated peripheral blood mononuclear cells from healthy donor were co-cultured with hPAMSCs supernatant for 6 hours.The percentage of CD4+T cells and Treg cells was detected by flow cytometry.ELISA was used to test the level of transforming growth factorβin the cell supernatant.RESULTS AND CONCLUSION:Successfully established primary hPAMSCs cell lines had stable passage.Supernatant of hPAMSCs significantly inhibited the percentage of CD4+T cells(P<0.05),while significantly enhanced the percentage of Treg cells(P<0.05).Meanwhile,the level of transforming growth factorβwas significantly up-regulated(P<0.01).In conclusion,the supernatant of hPAMSCs has the immunomodulatory function via inducing the conversion from CD4+T to Treg cells,which is expected to be a new therapeutic treatment for immune disorders.

关 键 词：胎盘间充质干细胞 CD4^+T细胞 Treg细胞 转化生长因子Β 免疫调节 干细胞 胎盘 间质干细胞 CD4阳性T淋巴细胞 T淋巴细胞 调节 转化生长因子Β 组织工程 

分 类 号：R459.9[医药卫生—治疗学]