体外诱导小鼠骨髓间充质干细胞分化为肝样细胞
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俞瑞珺, 刘海林
诱导分化;骨髓间充质干细胞;肝细胞;生长因子,俞瑞珺,刘海林,通讯作者:,Differentiationofmousebonemarrowmesenchymalstemcellsintohepatocyte-likecellsinvitro,Rui-Ju
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俞瑞珺, 刘海林, 上海交通大学医学院附属第九人民医院消化内科 上海市 200011
俞瑞珺, 2007届上海交通大学硕士, 内科住院医师, 主要从事肝硬化及肝细胞移植的研究.
通讯作者: 刘海林, 200011, 上海制造局路639号, 上海交通大学医学院附属第九人民医院消化内科. liuhailin@medmail.com.cn
电话: 021-53078068
收稿日期: 2007-07-16 修回日期: 2007-11-10
Differentiation of mouse bone marrow mesenchymal stem cells into hepatocyte-like cells in vitro
Rui-Jun Yu, Hai-Lin Liu
Rui-Jun Yu, Hai-Lin Liu, Department of Gastroenterology, the Ninth People’s Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200011, China
Correspondence to: Dr. Hai-Lin Liu, Department of Gastroenterology, the Ninth People’s Hospital, School of Medicine, Shanghai Jiaotong University, 639 Zhizaoju Road, Shanghai 200011, China. liuhailin@medmail.com.cn
Received: 2007-07-16 Revised: 2007-11-10
Abstract
AIM: To induce hepatocyte-like cells from mouse bone marrow mesenchymal stem cells, and improve their differentiation rate in vitro.
METHODS: Bone marrow mesenchymal stem cells were collected from C57 mice, and cultured in differentiation medium including HGF, FGF4, EGF, OSM, Dex and ITS. Expression of hepatocyte specific genes, such as AFP, Alb, CK18, TAT and CYP2b9 was detected by RT-PCR. Cells expressing Alb and CK18 proteins were identified by immunofluorescence staining and flow cytometry.
RESULTS: In the differentiation group, some polygonal cells appeared on day 3 in culture, cobblestone-shaped epithelial-like cells appeared after 5 to 7 days and increased in number until day 14, while control group cells retained their spindle morphology. AFP, Alb and CK18 mRNA could be detected on days 7, 14 and 21. TAT and CYP2b9 mRNA were also detected on days 14 and 21. However, Alb, CK18, TAT and CYP2b9 mRNAs were not detected in the control group, and only low levels of AFP mRNA were detected. In the differentiation group, Alb protein was detected by immunofluorescence on days 7, 14 and 21 ......
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