基础医学与临床 ›› 2012, Vol. 32 ›› Issue (9): 998-1003.

• 研究论文 • 上一篇    下一篇

人羊膜上皮细胞在大鼠受损肝组织中分化为肝细胞

宫黎明1,方宁1,陈代雄1,万卫红1,章涛1,余丽梅1,赵春华2   

  1. 1. 贵州省细胞工程重点实验室
    2. 中国医学科学院基础医学研究所组织工程中心
  • 收稿日期:2011-04-21 修回日期:2011-12-26 出版日期:2012-09-05 发布日期:2012-08-28
  • 通讯作者: 陈代雄 E-mail:cellgene@163.com
  • 基金资助:
    “重大新药创制”国家科技重大专项课题;贵州省科技计划发展项目

Differentiation of human amnion epithelial cells into hepatocytes in rat injured liver

  • Received:2011-04-21 Revised:2011-12-26 Online:2012-09-05 Published:2012-08-28

摘要: 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 采用胰蛋白酶消化法从羊膜组织中分离人羊膜上皮细胞(hAECs),采用流式细胞术和免疫荧光染色进行表型分析和细胞鉴定。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hAECs移植组和对照组,每组20只。造模后24h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48h、1周、2周和4周处死各实验组5只大鼠,取肝脏制备冰冻切片,采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:⑴ 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;⑵ 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。

关键词: 人羊膜上皮细胞, 移植, 分化, 肝细胞, 肝损伤

Abstract: Abstract:Objective To study the survival and differentiation into hepatocytes of hAECs in vivo. Methods hAECs 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, 50μl 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 ⑴Freshly isolated hAECs expressed CD29 and CD166; immunofluoresce staining showed that cytokeratin 19 was positive in hAECs, while vimentin was negative. ⑵hAECs transplanted into injured liver were located in hepatic lobules at 48h, and expressed AFP at 1w, CK18 at 2w, and Alb at 4w after transplantation. Conclusion hAECs xenografted to rat injured liver can differentiate into hepatocytes, suggesting that hAECs may have the potential for treating clinical liver injury diseases.

Key words: human amnion epithelial cell, transplantation, differentiation, hepatocyte, liver injury

中图分类号: