肝衰竭患者血浆置换前后血清诱导脐带间充质干细胞向肝细胞的分化  

Hepatic failure patient's serum before and after plasmapheresis induces the differentiation of umbilical cord mesenchymal stem cells into hepatic cells

作　　者：王晶波[1] 耿大影[2] 李晓迎[1] 陈莉 徐凤 史兆章[4] Wang Jing-bo Geng Da-ying Li Xiao-ying Chen Li Xu Feng Shi Zhao-zhang(Third Department Centra Laboratory 4Department of Intervention, Jinan Municipal Hospital for Infectious Diseases, Jinan 250021, Shandong Province, Chin 3Cell Biology, Jinan 250021, Shandong Province, Chin)

机构地区：[1]济南市传染病医院三科,山东省济南市250021 [2]济南市传染病医院中心实验室,山东省济南市250021 [3]济南赛尔生物科技有限公司,山东省济南市250021 [4]济南市传染病医院介入科,山东省济南市250021

出　　处：《中国组织工程研究》2017年第17期2659-2664,共6页Chinese Journal of Tissue Engineering Research

基　　金：2012年度济南市科技局无资计划项目;山东省医药卫生科技发展计划项目(2014WS0006)~~

摘　　要：背景:脐带间充质干细胞治疗肝衰竭已经取得了一定的疗效,但是肝衰竭患者的血清微环境对脐带间充质干细胞分化的影响尚未见报道。目的:探讨慢加急性肝衰竭患者血浆置换前、后血清对脐带间充质干细胞向功能肝细胞分化的影响。方法:采用组织块贴壁培养法获得脐带间充质干细胞并进行细胞形态及表型鉴定;取第3代脐带间充质干细胞,用含有肝衰竭患者血浆置换前、后血清的α-MEM培养基诱导培养,常规胎牛血清培养基为对照组,显微镜下观察各组细胞的形态,免疫组化检测细胞甲胎蛋白、白蛋白和细胞角蛋白18的表达水平。结果与结论:(1)组织块贴壁培养法能够获得大量高纯度的脐带间充质干细胞,细胞高表达CD44,CD73,CD90,CD105,不表达CD45,符合间充质干细胞的表型特征;(2)慢加急性肝衰竭患者血清可改变脐带间充质干细胞的形态,并诱导脐带间充质干细胞表达甲胎蛋白、白蛋白和细胞角蛋白18。血浆置换后血清组3种蛋白阳性率高于血浆置换前血清组,差异有显著性意义(P<0.05);(3)结果表明,肝衰竭患者血浆置换后血清更加有利于脐带间充质干细胞生长及向肝细胞分化。BACKGROUND： Umbilical cord mesenchymal stem cells （UC-MSCs） have made certain curative effect on hepatic failure, but little is reported on the effect of hepatic failure patient＇s serum microenvironment on UC-MSCs differentiation ability. OBJECTIVE： To investigate the effect of hepatic failure patient＇s serum on the differentiation of umbilical cord mesenchymal stem cells into hepatic cells. METHODS： UC-MSCs were isolated by tissue adherent method and the cell morphology and phenotype identified by microscope and flow cytometry. Alpha-MEM media with serum before/after plasmapheresis were used to culture the hirdgeneration of UC-MSCs, and regular fetal bovine serum culture medium acted as control group. Inverted microscope was used to observe the cell morphology in three groups. Immunohistochemical method was used to measure expression level of alpha fetoprotein, albumin and cytokeratin 18. RESULTS AND CONCLUSION： A great amount of high-purity UC-MSCs could be obtained using tissue adherent method, which highly expressed CD44, CD73, CD90, CD10, but did not express CD45. Hepatic failure patient＇s serum could change the morphology of UC-MSCs and induce UC-MSCs to express alpha fetoprotein, albumin and cytokeratin 18. The positive expression of alpha fetoprotein, albumin and cytokeratin 18 was significantly increased after plasmapheresis （P 〈 0.05）. To conclude, hepatic failure patient＇s serum after plasmapheresis exert more benefits to induce UC-MSCs to differentiate into hepatic cells than that before plasmapheresis.

关 键 词：干细胞 脐带脐血干细胞 脐带间充质干细胞 血浆置换 肝衰竭 肝细胞分化 

分 类 号：R394.2[医药卫生—医学遗传学]