人羊膜间充质干细胞在大鼠受损肝组织中分化为肝细胞  

Differentiation of human amnion derived-mesenchymal stem cells into hepatocytes in rat injured liver

作　　者：宫黎明[1] 方宁[1] 陈代雄[1] 万卫红[1] 章涛[1] 刘祖林[1] 余丽梅[1] 赵春华[2] 

机构地区：[1]遵义医学院附属医院贵州省细胞工程重点实验室,贵州省遵义市563003 [2]中国医学科学院基础医学研究所组织工程研究中心,北京市100005

出　　处：《中国组织工程研究与临床康复》2011年第36期6709-6713,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金："重大新药创制"国家科技重大专项课题(2009ZX09503-025);贵州省科技计划发展项目(黔科合计字2051号;黔科合SZ字3017号)~~

摘　　要：背景:羊膜间充质干细胞在体外适当的诱导条件下可分化为肝细胞样细胞,而在受损肝脏原位是否可分化为肝细胞值得探讨。目的:观察人羊膜间充质干细胞在大鼠肝损伤原位植活及分化情况。方法:采用胰酶-胶原酶二酶消化法从羊膜组织中分离人羊膜间充质干细胞。采用腹腔注射D-氨基半乳糖建立大鼠肝损伤模型,随机分为2组,人羊膜间充质干细胞移植组造模后注射L-DMEM悬浮的人羊膜间充质干细胞悬液,对照组注射等量L-DMEM。结果与结论:①FCM分析结果显示,所分离的人羊膜间充质干细胞表达CD29、CD44和CD166;免疫荧光染色显示人羊膜间充质干细胞表达波形蛋白,不表达CK19。②与对照组比较,人羊膜间充质干细胞移植后肝病理无明显差异,均呈急性肝坏死病理改变。③免疫荧光双染色结果显示,人羊膜间充质干细胞移植后1周主要定植于肝小叶且表达CK19,至2周表达CK18,至3周表达Alb。结果表明,人羊膜间充质干细胞在大鼠受损肝组织中能被植活,且可分化为肝细胞,提示人羊膜间充质干细胞移植在临床肝病的治疗方面可能具有潜在应用价值。BACKGROUND：Human amnion derived-mesenchymal stem cells（hAD-MSCs） can differentiate into hepatocyte-like cells in certain conditions in vitro.It needs to further investigate whether they can differentiate into hepatocytes in damaged liver.OBJECTIVE：To study the survival and differentiation of hAD-MSCs in vivo in rat injured liver.METHODS：hAD-MSCs were isolated from human amnion treated with trypsin and collagenase.Forty healthy and clean grade female SD rats underwent intraperitoneal injection of D-galactosamine diluted in normal saline at the dose of 400 mg/kg body weight,to establish the model of liver injury,and then divided into hAD-MSCs group and control group stochastically.Twenty-four hours after modeling,50 μL cell suspension（approximately 1×10^6 cells suspended in L-DMEM） of hAD-MSCs were injected slowly into the left,middle and right hepatic lobe respectively with micro-syringe,while equivalent volume of L-DMEM injected into the control group.RESULTS AND CONCLUSION：①Freshly isolated hAD-MSCs expressed CD29,CD44 and CD166;immunofluorescence staining showed that vimentin was positive in hAD-MSCs,while cytokeratin 19 was negative.②There was no significant difference after transplanting hAD-MSCs into injured liver between the hAD-MSCs group and the control group,remaining acute hepatic necrosis.③hAD-MSCs implanted into injured liver were located in hepatic lobules at day 7 after transplantation,which expressed CK19;and the cells expressed CK18 at day 14,and also human Alb at day 21.hAD-MSCs xenografted to rat injured liver can survive and differentiate into hepatocytes,suggesting that hAD-MSCs transplantation may have potential applications for treating clinical liver injury diseases.

关 键 词：人羊膜间充质干细胞 移植 分化 肝细胞 肝损伤 

分 类 号：R394.2[医药卫生—医学遗传学]