人羊膜上皮细胞对四氯化碳诱导小鼠肝损伤的定位修复  

Human amnion epithelial cell transplantation provides a local repair of liver damage induced by carbon tetrachloride in mice

作　　者：刘默凝 张钰堃 李岩[1] 曹贵方[1] 白立恒 Liu Mo-ning;Zhang Yu-kun;Li Yan;Cao Gui-fang;Bai Li-heng(Laboratory of Animal Embryology and Developmental Biology,Inner Mongolia Agricultural University,Hohhot 010018,Inner Mongolia Autonomous Region,China;Department of Maternity,Inner Mongolia Maternal and Child Health Hospital,Hohhot 010020,Inner Mongolia Autonomous Region,China)

机构地区：[1]内蒙古农业大学动物组织胚胎学与发育生物学实验室,内蒙古自治区呼和浩特市010018 [2]内蒙古妇幼保健医院,内蒙古自治区呼和浩特市010020

出　　处：《中国组织工程研究》2018年第5期698-703,共6页Chinese Journal of Tissue Engineering Research

基　　金：内蒙古自治区自然科学基金项目(2015MS0375)~~

摘　　要：背景:人羊膜上皮细胞具有无致瘤性、来源丰富、容易获取、免疫原性低、无医学伦理限制等优点,很多优势是胚胎干细胞和诸多成体干细胞所不具备的,有望成为今后临床移植治疗的重要种子细胞。目的:观察并研究荧光染料羟基荧光素二醋酸盐琥珀酰亚胺脂(5,6-carboxyfluorescein diacetate succinimidyl ester,CFSE)标记的人羊膜上皮细胞对四氯化碳诱导的小鼠肝损伤的定位修复情况。方法:使用酶消化法从人羊膜中分离得到人羊膜上皮细胞,观察细胞形态,人羊膜上皮细胞的鉴定采用免疫细胞化学法检测细胞角蛋白19的表达。用四氯化碳腹腔注射诱导小鼠肝损伤。将用CFSE标记的人羊膜上皮细胞通过尾静脉注射移植到肝损伤小鼠体内,细胞移植后7 d后检测小鼠组织病理学变化,30 d后检测肝功相关指标。结果与结论:①实验成功分离得到纯度较高的人羊膜上皮细胞;②免疫细胞化学检测显示人羊膜上皮细胞可表达上皮细胞标志角蛋白19;③冰冻切片免疫荧光显示人羊膜上皮细胞可以归巢到受损肝脏;④人羊膜上皮细胞移植后对肝损伤小鼠的肝功能、组织病理学改变都有明显修复作用; ⑤结果证实,人羊膜上皮细胞可有效改善肝组织的生理功能,为细胞定位移植治疗肝脏疾病的修复情况提供实验数据。BACKGROUND:Human amnion epithelial cells(hAECs)have many merits that embryonic stem cells and adult stem cells do not have,such as no tumorigenicity,rich sources,easy to obtain,low immunogenicity and no medical ethics limit.Therefore,hAECs are expected to be important seed cells for clinical transplantation.OBJECTIVE:To observe the therapeutic effect of hAECs transplantation labeled with carboxyfluorescein diacetate succinimidyl ester(CFSE)in a mouse model of liver damage induced by carbon tetrachloride.METHODS:hAECs from the human amniotic membrane were collected using enzymatic digestion,and morphology of cells was observed.Expression of keratin 19 in hAECs was detected by immunocytochemistry.Model of liver damage was made in mice by intraperitoneal injection of carbon tetrachloride.Then,CFSE-labeled hAECs were injected into the liver damage mice via tail vein.Histopathological changes and liver function in mice were observed at 7 and 30 days after transplantation,respectively.RESULTS AND CONCLUSION:The high-purity hAECs were successfully isolated,which expressed keratin 19 shown by immunocytochemical staining.Frozen sections of immunoflrorescence showed that hAECs could be moved to the damaged liver,and exhibited remarkable repair effects on the liver function and histopathology in mice.These findings indicate that hAECs can be used for xenotransplantation and function to promote physiological recovery from liver injury,thereby providing experimental evidence for liver repair with cell transplantation.

关 键 词：人羊膜上皮细胞 四氯化碳 肝损伤 定位修复 CFSE 谷草转氨酶 冰冻切片 干细胞 免疫荧光 小鼠 组织工程 

分 类 号：R394.2[医药卫生—医学遗传学]