【摘要】 目的探讨人羊膜上皮细胞在大鼠损伤肝原位植活及向肝细胞分化。方法用胰蛋白酶消化法从羊膜组织中分离人羊膜上皮细胞(hAECs),用流式细胞术和免疫荧光染色进行表型分析和细胞鉴定。腹腔注射D-氨基半乳糖溶液建立大鼠肝损伤模型,随机分为hAECs移植组和对照组,每组20只。造模后24 h用微量注射器于肝左、中、右叶3点分别移植L-DMEM悬浮的hAECs悬液50μL(约1×106个细胞),对照组注射等量L-DMEM。于移植后48 h、1、2和4周处死各实验组5只大鼠,取肝脏制备冰冻切片,用免疫荧光双染色检查hAECs在受损肝原位的植活、分布及其向肝细胞分化的标志物表达。结果 1)FCM分析和免疫荧光染色结果显示,所分离的hAECs表达CD29、CD166及CK19,几乎不表达CD44、CD80、CD86、HLA-DR及波形蛋白;2)免疫荧光双染色结果显示,hAECs移植后1周主要定植于肝小叶且表达AFP,至2周表达CK18,至4周表达Alb。结论 hAECs在大鼠受损肝组织中能被植活且可分化为肝细胞,提示hAECs移植在临床肝病的治疗方面可能具有潜在应用价值。更多还原

【Abstract】 Objective To investigate the survival and differentiation into hepatocytes of hAECs in rat.MethodshAECs were isolated from human amnion treated with trypsin,and the phenotype and characteristics of immunocytochemistry were analyzed by FCM and immunofluorescence staining.Healthy and clean grade female SD rats were administered intraperitoneal injection of D-galactosamine diluted in normal saline at a dose of 400 mg/kg body weight,to establish liver injury model.Rats were then devided into hAECs group and control group stochastically,with twenty in each group.Twenty-four hours after modeling,50 μL cell suspension(approximately 1×106 cells suspended in L-DMEM) of hAECs was injected slowly into the left,middle and right lobe respectively with micro-syringe,while equivalent volume of L-DMEM injected into the control group.Results 1)Freshly isolated hAECs expressed CD29 and CD166;immunofluoresce staining showed that cytokeratin 19 was positive in hAECs,while vimentin was negative.2)hAECs transplanted into injured liver were located in hepatic lobules at 48 h,and expressed AFP at 1w,CK18 at 2w,and Alb at 4w after transplantation.Conclusions hAECs xenografted to rat injured liver can differentiate into hepatocytes suggesting hAECs may have the potential value for treating clinical liver injury.更多还原