小鼠胚胎干细胞诱导为肝细胞的研究  

Study of Hepatocytes Induced by Cultured Mouse Embryonic Stem Cells

作　　者：蒯小玲[1] 萧树东[1] 丛笑倩[2] 李秀兰[2] 

机构地区：[1]上海第二医科大学附属仁济医院上海市消化疾病研究所,200001 [2]中国科学院上海细胞生物研究所

出　　处：《胃肠病学》2003年第1期6-10,共5页Chinese Journal of Gastroenterology

基　　金：本课题由上海市重点学科经费资助

摘　　要：背景:胚胎干细胞(ES细胞)是从体外受精胚囊的内细胞团分离建立的、具有发育全能性的细胞系,在特定条件下可向多种细胞分化,是细胞移植治疗很有前途的细胞来源。目的:探明ES细胞是否能被诱导分化为肝细胞,并探索小鼠ES细胞诱导分化为肝细胞的分化条件。方法:在ES细胞培养液中分别加入肝细胞生长因子(HGF)、β-神经细胞生长因子(NGF)和维甲酸(RA)以及与小鼠胎肝细胞共培养,观察分化细胞的形态学变化,逆转录聚合酶链反应(RT-PCR)检测白蛋白和转甲状腺蛋白mRNA水平的表达,免疫组化法检测甲胎蛋白和α1-抗胰蛋白酶蛋白水平的表达。结果:ES细胞培养液中加入HGF和β-NGF 15天后,分化出较多的上皮样细胞,并检测出白蛋白、转甲状腺蛋白mRNA水平的表达和甲胎蛋白、α1-抗胰蛋白酶蛋白水平的表达,表明ES细胞己诱导分化为肝细胞。ES细胞与胎肝细胞共培养2天后,分化出单一形态的上皮细胞,同样有上述肝细胞标志物的阳性表达。RA诱导出的细胞除转甲状腺蛋白mRNA外,无其他肝细胞标志物的阳性表达。结论:在HGF和β-NGF的作用下,有部分小鼠ES细胞被诱导为肝细胞,RA则无此作用。共培养方法也能诱导出肝细胞标志物阳性的细胞,并且细胞形态较单一。小鼠ES细胞有向肝细胞分化的潜能。Background: Embryonic stem (RS) cells are pluripotent cells derived from the inner cell mass of in vitro fertilized blastocysts. It can be induced to undergo differentiation into all cell types in specific conditions and may be used as the cell resource of transplantation. Aims: To explore whether the cultured mouse ES cells can be induced into hepatocytes and the conditions of differentiation. Methods: After hepatocyte growth factor (HGF), p-nerve growth factor (NGF) and retinoic acid (RA) were added into cell culture respectively, and co-culture with mouse embryonic hepatocytes, the morphological changes of the mouse ES cells were observed; albumin and transthyretin mRNA were examined by reverse transcriptase polymerase chain reaction (RT-PCR), and al-antitrypsin and x-fetoprotein were assayed by immunohistochemistry. Results: After the addition of HGF and x-NGF and co-culture with mouse embryonic hepatocytes, albumin and transthyretin mRNA were detected by RT-PCR, al-anti-trypsin and cx-fetoprotein were detected by immunohistochemistry in some differentiated cells. But RA had no such response except that transthyretin mRNA was positive. Conclusions: Mouse ES cells can be induced into hepatocyte-like cells by HGF and P-NGF as well as co-culture with mouse embryonic hepatocytes, whereas RA has no such effect. Mouse ES cells have the potential to be directed into hepatocytes.

关 键 词：肝细胞 胚胎干细胞 体外受精 胚囊 细胞培养液 肝细胞生长因子 HGF β-神经细胞生长因子 NGF 维甲酸 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]