体外三维培养骨髓间充质干细胞诱导分化为肝细胞及其极化特征

基础医学与临床 ›› 2012, Vol. 32 ›› Issue (6): 639-643.

体外三维培养骨髓间充质干细胞诱导分化为肝细胞及其极化特征

卫金花¹,曲强²,杨阳¹,何小东³,刘卫⁴

1. 中国医学科学院北京协和医学院北京协和医院
2. 中国医学科学院北京协和医学院北京协和医院
3. 北京协和医院
4. 中国医学科学院北京协和医院

收稿日期:2012-04-05 修回日期:2012-04-25 出版日期:2012-06-05 发布日期:2012-05-25
通讯作者: 曲强 E-mail:qiangqu@hotmail.com

Differentiated hepatocytes derived from human bone mesenchymal stem cell in 3-dimentional culture and the polarity characteristic

Received:2012-04-05 Revised:2012-04-25 Online:2012-06-05 Published:2012-05-25

摘要/Abstract

摘要： 摘要：目的探讨体外三维诱导培养体系下人骨髓间充质干细胞向肝细胞分化及极化特征。方法从成人骨髓中分离出骨髓间充质干细胞(Bone mesenchymal stem cell, BMSC),利用胶原蛋白三维培养体系进行体外诱导培养.用HE染色法观察分化肝细胞的形态及结构特征,用免疫组化法检测肝细胞特异性蛋白的表达,用免疫荧光法检测极化蛋白BSEP和SRBI的表达特征。结果成人BMSC在体外胶原蛋白三维诱导培养体系中呈多层排列生长,形成细胞间紧密连接及管状结构。BMSC分化来源肝细胞特异性表达ALB及AFP。肝细胞极化蛋白BSEP和SRBI在肝细胞呈特征性分布。结论 BMSC在体外胶原三维培养体系中可诱导分化为肝细胞,并形成肝组织特征性的极化结构。

关键词: 骨髓间充质干细胞, 三维培养, 肝细胞, 极化, 分化

Abstract: Abstract: Objective To explore hepatic differentiation of human bone mesenchymal stem cell (BMSC) in 3-dimentional (3D) culture system of rat type-I collage and the polarity of differentiated hepatocyte. Methods BMSCs were isolated from adult bone marrow and differentiated to hepatocyte in 3D culture system of rat type-I collage. The morphological and structural characteristics of differentiated hepatocyte derived from BMSC in 3D culture were observed using HE staining. The expression of hepatocyte-specific markers of ALB and AFP were analyzed using immunohistochemical staining. The expression of hepatocyte polarity protein BSEP and SRB1 were assessed using the Immunofluorescence. Results Differentiated hepatocyte derived from BMSC formed tight junction and tubular or spherical structures in 3D culture system. The expression of ALB and AFP in hepatocytes differentiated from BMSC was positive. The expression of BSEP and SRBI in the 3D culture was positive and characteristic. Conclusion BMSC can differentiate to hepatocyte in 3D culture. The differentiated hepatocyte can form hepatocyte-specific polarity in 3D culture.

Key words: Bone mesenchymal stem cell, 3-dimentional culture, Hepatocyte, Polarity, Differentiation

中图分类号:

R318.14

卫金花曲强杨阳何小东刘卫. 体外三维培养骨髓间充质干细胞诱导分化为肝细胞及其极化特征[J]. 基础医学与临床, 2012, 32(6): 639-643.

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