文献类型：期刊文献

中文题名：体外分步诱导猕猴胚胎干细胞为肝细胞

英文题名：Induction of Rhesus Monkey Embryonic Stem Cells into Hepatocyte-like Cells by a Three-step Method

作者：金立方[1,2];纪少珲[2];郭祥玉[2];王喜宏[2];季维智[2]

机构：[1]绍兴文理学院生命科学学院;[2]中国科学院昆明动物研究所,云南省动物生殖生物学重点实验室

年份：2009

卷号：30

期号：5

起止页码：509

中文期刊名：动物学研究

外文期刊名：Zoological Research

收录：CSTPCD、、Scopus、CSCD2011_2012、北大核心2008、北大核心、CSCD、PubMed

基金：973计划(2006CB701505;2007CB947701);中科院创新工程试点经费(KSCX2-YW-R-47);国家自然科学基金(30570906;30700159;30700425)

语种：中文

中文关键词：猕猴;胚胎干细胞;肝细胞;细胞因子

外文关键词：Rhesus monkey; Embryonic stem cells; Hepatocyte-like cells; Growth factors

中文摘要：采用分阶段诱导方法模拟肝细胞体内发育,建立体外诱导猕猴胚胎干细胞(rhesusmonkeyembryonicstemcells,rESCs)分化为成熟肝细胞的体系,对研究以ES细胞为基础的临床替代治疗人类晚期肝脏疾病具有重要的意义。将rESCs团块在含有10%FBS的DMEM培养基中悬浮培养11d,形成含有早期内胚层细胞的拟胚体(embryonicbodies,EB)并开始表达早期肝细胞的部分基因或蛋白,将11日龄EB接种至包被有ECM的组织培养皿,分阶段加入aFGF、BMP-4及OSM。经aFGF和BMP-4诱导7～10d后,分化细胞形态变为具有双核的多角形细胞,表达早期和中期肝细胞特异性的蛋白(AFP、ALB及CK18)和基因(AFP、ALB、APOH,G-6-P及TAT),并具有储存糖原的功能。撤除aFGF和BMP-4,添加OSM继续诱导7～10d,分化的细胞表达成熟肝细胞所特有基因CYP1B1和ADH1C,并具有摄取靛青绿的能力。

外文摘要：Embryonic stem cells （ESCs） hold great potential for the treatment of liver diseases. Here, we reported the induction of rhesus monkey ESCs along a hepatocyte lineage by a three-step method. The Rhesus monkey ESCs were suspended in DMEM medium supplemented with 10% FBS for 11 days to form embryonic bodies （EB） which contained early endoderm or hepatic cells by expression of early hepatic genes. EB of 11 days were transferred into ECM coated dishes and cultured with aFGF, BMP-4 and OSM at different time points for further studies. After induction by aFGF and BMP-4 for 7 - 10 days, the differentiated cells showed a hepatocyte-like morphology, expressed hepatic marker genes （such as AFP, ALB APOH, G-6-P and TAT） and proteins （AFP, ALB and CK18）, and possessed the capacity to storage intracytoplasmic glycogen. After further induction with OSM for 7 - 10 days, some cells shared similar characteristics of matured hepatocytes, such as expression of genes of CYP1B1 and ADHIC, and ICG uptaken, indicating the presence of functional hepatocytes in the differentiated cells.