多次移植人脐带间充质干细胞对大鼠Th1和Th2比值的影响  

The effects of xenotransplantation of human umbilical cord mesenchymal stem cell on the Th1/Th2 of the rat

作　　者：李娜[1] 辛毅[1] 林筝[1] 孔晴宇[2] 张颖[1] 周玉杰[3] 黄益民[1] 张兆光[2] 

机构地区：[1]首都医科大学附属北京安贞医院-北京市心肺血管疾病研究所细胞与分子生物学研究室,北京100029 [2]首都医科大学附属北京安贞医院-北京市心肺血管疾病研究所心外科,北京100029 [3]首都医科大学附属北京安贞医院-北京市心肺血管疾病研究所心内科,北京100029

出　　处：《心肺血管病杂志》2013年第6期789-793,共5页Journal of Cardiovascular and Pulmonary Diseases

基　　金：国家自然科学基金(No.81370225);北京市优秀人才培养资助项目(NO:2012D003034000009);北京市自然科学基金支持项目(No.5042006;7072012)

摘　　要：目的：观察多次静脉移植人脐带间充质干细胞（human umbilical mesenchymal stem cells,HUMSCs）对大鼠Th1和Th2轴的影响.方法：①体外培养HUMSCs;流式细胞仪鉴定第4代hUMSCs表面抗原的表达；特异性诱导染色并鉴定第4代HUMSCs分化脂肪细胞和成骨细胞的潜能；②雄性SD大鼠30只,体质量182～203g,分为3组：空白对照组；0.9％氯化钠液组；干细胞移植组,每组10只.干细胞移植组：1次/w经尾静脉注射3.5×107/kg的HUMSCs,连续3次.第三次移植HUMSCs后11d,各组经腹主动脉取血,流式细胞仪检测T淋巴细胞的IFN-γ和IL-4阳性率.结果：3次注射干细胞后实验动物全部存活,干细胞组的Th1、Th2阳性率较0.9％氯化钠液组差异无统计学意义（P＞0.05）；Th1/Th2比值、血清IFN-γ和IL-4水平与对照组比较差异无统计学意义（P＞0.05）；干细胞组的心、脑、肺、肝及肾等脏器未发生病理改变.利用红色荧光蛋白转染的HUMSCs示踪,发现移植的HUMSCs大部分归巢于骨髓中.结论：多次尾静脉注射HUMSCs,未引起大鼠Th1、Th2细胞免疫和体液免疫系统的变化,Th1/Th2轴无明显改变.移植的HUMSCs能较长时间的存活于大鼠骨髓内,具有较好的免疫相容性.Objective：To investigate the effects of multiple circles exotransplantations of Human um- bilical cord mesenchymal stem cell （HUMSCs） on the T helper cell relative immunology system in SD rats. Method： HUMSCs derived from Wharton＇s jelly tissue were aseptically isolated and expanded in vitro. 3.5 × 10^7/kg HUMSCs were transplanted to the SD rats through the tail vein once a week for a total of 4 weeks. The saline solution group served as the control group. The blood samples were drew through the abdominal aorta one day after the last transplantation. The lymphocytes were isolated and stained by the CIM, CD8, I1-4 and IFN-~/ fluorescent antibody. The Th1 ,Th2 percentage were tested by flow cytometry. The brain, heart, lung, liver and kidney were collected for HE staining. Results ： （1） the 4th passage of the HUMSCs expressed positively the spe- cific surface antigen such as CD73,CD90 and CD105, and the positivity arrived 99.4%, 100% and 99.9% ; The expression of CD34, CIM5, CDllb, CD19, CD79a and HLA-DR were negative. The differentiation rate of the HUMSCs towards the osteoblasts and adipocytes 98.4% and 95.7% ;（2）experimental animals all survived after HUMSCs treatment, the positive percentage of Thl （ IL-4 ＋ cells） and Th2 （ IFN-γ ＋ cells） did not showany significant difference compare to the control. And the Thl/Th2 axis did not changed significantly by the HUMSCs transplantation [ （2. 59 ± 1.42） vs. （2. 34 ± 0. 58）, P 〉 0. 05 ]. （3） HE staining showed that no sig- nificantly morphological and pathological changes in the brain, heart, lung, liver and the kidney in the HUM- SCs groups. Conclusion ： The different dose of HUMSCs intravenous infusion of the HUMSCs doesn＇t have sig- nificant effects on the Tbl, Th2 and the Thl/Th2 percentage. And the transplanted HUMSCs survive majorly in the bone marrow for 4 weeks after the transplantation. The xenotransplantation of HUMSCs in the rats is biocom- patible and did not affect the rats immune system.

关 键 词：人脐带胶样组织间充质干细胞 异种移植 免疫反应 动物实验 大鼠 

分 类 号：R446.7[医药卫生—诊断学]