人脐血间充质干细胞抑制异体T淋巴细胞反应的实验及临床意义.doc

人脐血间充质干细胞抑制异体T淋巴细胞反应的实验及临床意义
作者:王茜,杨琨,白海, 吴涛, 路继红
【摘要】目的: 研究人脐血间充质干细胞(MSCs)对异体外周血T淋巴细胞抑制的影响. 方法:分离、扩增脐血MSCs,流式细胞术检测表面标志以鉴定. 取分离培养的脐血MSCs,与经分离的异体淋巴细胞共培养,在植物血凝素(PHA)刺激下,用MTT还原法测定细胞增殖,观察脐血MSCs对PHA刺激的T淋巴细胞转化抑制作用. 采用ELISA检测培养上清中IFNγ和IL4的分泌水平. 结果:脐血MSCs在PHA刺激下可抑制T淋巴细胞增殖,且其培养上清也具有抑制异体淋巴细胞增殖转化能力. ELISA检测7 d共培养上清中IFNγ和IL4两种细胞因子的含量,发现IFNγ分泌水平下调、IL4分泌水平部分上调. 结论:脐血MSCs具有抑制异体淋巴细胞免疫反应,降低移植物抗宿主反应(GVHD)的作用. 具有研究价值和应用潜能.
【关键词】胎血/细胞学;间质干细胞;外周血T淋巴细胞;植物血凝素类;干扰素Ⅱ型;白细胞介素4
【Abstract】 AIM: To study the effects of human umbilical cord bloodderived mesenchymal stem cells (MSCs) on peripheral blood T lymphocytes. METHODS: MSCs were isolated from normal human umbilical cord blood, cultured in vitro, purified and then identi
fied by examining surface marker with FCM. Lymphocytes were isolated from allogenic peripheral blood and identified with FCM. The MSCs and lymphocytes were cocultured in 96well plates for 72 h. T lymphocyte proliferation was measured by using MTT reduction method and the effect of MSCs on the T lymphocyte transformation stimulated by PHA was observed. On day 7 of coculture, the culture supernatant was collected to detect the expressions of IFNγ and IL4 with ELISA. RESULTS: Human umbilical cord bloodderived MSCs inhibited T lymphocyte proliferation and PHA induced lymphocyte transformation. The secretion of IFNγ in supernatant was significantly downregulated,w