中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (14): 2491-2494.doi: 10.3969/j.issn.1673-8225.2011.14.005

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

骨髓间充质干细胞向肝样细胞的诱导及分化

范敬静1，李东良1，何秀华2，程变巧1，马  明1，张志强1   

  1. 1福建医科大学福总临床医学院肝胆内科，福建省福州市  350025
    2泉州市第一医院感染科，福建省泉州市  362000
  • 收稿日期:2011-01-10 修回日期:2011-02-16 出版日期:2011-04-02 发布日期:2013-11-02
  • 通讯作者: 李东良，教授，硕士生导师，福建医科大学福总临床医学院肝胆内科，福建省福州市350025 dongliangli@gmail.com.cn
  • 作者简介:范敬静★，女，1985年生，河北省赤城县人，汉族，福建医科大学福总临床医学院在读硕士，主要从事器官移植相关肝病的研究。 duimantian@126.com

Induction and differentiation of bone marrow mesenchymal stem cells into hepatocyte-like cells

Fan Jing-jing1, Li Dong-liang1, He Xiu-hua2, Cheng Bian-qiao1, Ma Ming1, Zhang Zhi-qiang1   

  1. 1Department of Hepatobiliary Medicine, Clinical Medical College of Fuzhou General Hospital, Fujian Medical University, Fuzhou  350025, Fujian Province, China
    2Department of Infectious Disease, First Hospital of Quanzhou, Quanzhou  362000, Fujian Province, China
  • Received:2011-01-10 Revised:2011-02-16 Online:2011-04-02 Published:2013-11-02
  • Contact: Li Dong-liang, Professor, Master’s supervisor, Department of Hepatobiliary Medicine, Clinical Medical College of Fuzhou General Hospital, Fujian Medical University, Fuzhou 350025, Fujian Province, China dongliangli@gmail.com.cn
  • About author:Fan Jing-jing★, Studying for master’s degree, Department of Hepatobiliary Medicine, Clinical Medical College of Fuzhou General Hospital, Fujian Medical University, Fuzhou 350025, Fujian Province, China duimantian@126.com

PDF

437

被引次数

17

摘要：

背景：研究表明，在体外提供肝细胞及成纤维细胞生长因子等多种因子可诱导骨髓间充质干细胞向肝样细胞分化，但对其诱导的最佳环境还处于摸索阶段。
目的：观察体外应用肝细胞生长因子和成纤维生长因子4等多种因子联合诱导骨髓间充质干细胞向肝样细胞的分化能力。
方法：采用全骨髓贴壁培养法分离、扩增大鼠骨髓间充质干细胞，培养第3代时用肝细胞生长因子、成纤维生长因子4、胰岛素铁硒传递蛋白及地塞米松等联合诱导其向肝样细胞的分化。于诱导7，14，21 d后观察细胞形态变化，RT-PCR检测细胞白蛋白、甲胎蛋白、细胞角蛋白的mRNA表达；于诱导14，21 d行PAS糖原染色检测，并采用免疫细胞荧光法检测细胞白蛋白表达。
结果与结论：随着诱导时间的延长，骨髓间充质干细胞表现为肝细胞样，并呈集落生长，肝细胞特异性标志物逐渐出现和成熟。细胞诱导至7 d出现甲胎蛋白mRNA表达，14 d表达增高，21 d时表达下降；14 d时细胞角蛋白18、白蛋白 mRNA和糖原出现表达，并随时间延长表达逐渐增加。细胞诱导14 d时，胞浆白蛋白出现表达，至21 d时表达增强。证实骨髓间充质干细胞在肝细胞生长因子、成纤维生长因子4等多种因子诱导作用下，具有强大的向肝细胞分化能力。

关键词: 肝样细胞, 骨髓间充质干细胞, 细胞分化, 肝细胞生长因子, 成纤维生长因子4

Abstract:

BACKGROUND: The research has demonstrated that hepatocyte and fibroblast growth factor and other factors can induce bone marrow mesenchymal stem cells (BMSCs) into hepatocyte-like cells in vitro, but the optimum environment of induction is still in the exploratory stage.
OBJECTIVE: To observe the differentiation ability of BMSCs into hepatocyte-like cells by hepatocyte growth factor (HGF) and fibroblast growth factor 4 (FGF-4) and other factors.
METHODS: BMSCs of rats were isolated and proliferated by the method of whole bone marrow attachment culture. At the third passage, BMSCs were differentiated into hepatocyte-like cells by HGF, FGF-4, insulin iron-selenium transducin, and hexadecadrol in vitro. The morphological changes of cells were observed at 7, 14, and 21 days. The mRNA expression of albumin (ALB), alpha-fetoprotein (AFP) and cytokeratin 18 (CK 18) was detected by reverse transcription-polymerase chain reaction (RT-PCR). Glycogen was tested by periodic acid-schiff (PAS) at 14 and 21 days after induction, and the expression of ALB was detected by immunofluorescence method.
RESULTS AND CONCLUSION: With the prolonged induction time, BMSCs showed hepatocyte-like, round cells appeared and showed colony growth, liver cell-specific markers gradually emerge and mature. The AFP mRNA expression was appeared at 7 days after induction, and increased significantly at 14 days, then decreased at 21 days. Cells expressed CK18, ALB mRNA and glycogen at 14 days, and increased with the prolonged induction time. The expression of ALB in the cytoplasm is positive at 14 days after induction, and increased significantly at 21 days. BMSCs have the ability of differentiating into hepatocyte-like cells by the method of HGF, FGF-4 and other factors.

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