SD大鼠骨髓间充质干细胞-肝细胞共培养上清液细胞因子分析  

Cell factor analysis of SD rat BMSC-liver cells co-culture supernatant

作　　者：霍大浪[1] 刘代顺[1] 宁家辉[1] 张袁媛[1] 冯成军[1] 王娜[1] 

机构地区：[1]遵义医学院第三附属医院感染科,贵州遵义563000

出　　处：《重庆医学》2014年第28期3766-3768,共3页Chongqing medicine

基　　金：遵义市科技局联合基金[遵市科合社字(2011)14号]

摘　　要：目的检测SD大鼠骨髓间充质干细胞(BMSC)-肝细胞共培养上清液中的细胞因子,探讨细胞因子的作用,为生物人工肝及临床治疗肝功能衰竭提供理论依据。方法分离SD大鼠BMSC及原代肝细胞,将生长良好的第3代BMSC与肝细胞按1∶10比例体外共培养(实验组),以单独肝细胞(对照组)和单独BMSC(BMSC组)作为对照。观察不同情况下肝细胞生存时间和形态,培养48h后用RayBiotech抗体芯片检测细胞培养上清液中的细胞因子。结果实验组的肝细胞生长增殖迅速,肝细胞可存活14d;对照组的肝细胞生长增殖缓慢,细胞培养可存活9d。实验组上清液中的细胞因子表达谱发生了明显改变:IL-1、IL-6、IL-10均较对照组及BMSC组升高2倍以上,差异有统计学意义(P<0.01)。结论 BMSC-肝细胞共培养,BMSC可能通过分泌IL-1、IL-6和IL-10促进肝细胞生长,延长肝细胞生存时间。Objective To detect supernatant of SD rat bone marrow mesenchymal stem cells（BMSC）and liver cells,and explore the role of cytokines,and provide a theoretical basis for the biological artificial liver and clinical treatment of liver failure.MethodsBMSCs and primary hepatocytes of SD rat were isolated.BMSCs with its third generation and liver cells by the ratio of 1∶10were co-cultured.Hepatocytes and BMSC were treated as control group.Observation the cells′survival and morphology and analysis of cytokine after 48 hwere made by RayBiotech.Results BMSC-hepatocyte in the co-culture group grow and proliferated rapidly,and hepatocyte could survive up to 14days;while hepatocyte cultured alone,it grow slowly and survived only 9days.Expression of BMSC-liver cells supernatant changed obviously：interleukin 1,interleukin 6,interleukin 10 were higher than those in the control group,above 2times,which showed significant difference（P〈0.01）.Conclusion Co-culture of BMSC-liver cell could induce BMSC secrete paracrine and autocrine：IL-1and IL-6and IL-10,which could promote the growth of hepatocyte and extend the liver cell survival time.In-vitro BMSC-liver cells could provide biological artificial liver cell sources,as well as provide theory basis for using cytokine treatment for liver failure.

关 键 词：干细胞 骨髓 肝细胞 共同培养技术 细胞因子 

分 类 号：R363[医药卫生—病理学]