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[碱性成纤维细胞生长因子真核表达载体的构建及对兔骨髓基质细胞的转染]

[Construction of eukaryotic expression vectors of basic fibroblast growth factor and transfection in rabbit bone marrow stromal cells].

作者信息

Xu Zhong-Shi, Xiao De-Ming, Lin Bo-Wen, Lu Xiao-Hu, Li Ran

机构信息

Department of Orthopedics, Shenzhen People's Hospital, Shenzhen, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2009 Mar;29(3):446-9.

Abstract

OBJECTIVE

To investigate approach and possibility of transferring basic fibroblast growth factor (bFGF) gene into rabbit bone marrow stromal cells (BMSCs).

METHODS

The eukaryotic expression vectors harboring bFGF cDNA were constructed and transfected into rabbit BMSCs mediated by liposome. The transcription and expression of bFGF gene in the transfected BMSCs were detected by means of morphological observation, immunohistochemistry, enzyme-linked immunosorbent assay (ELISA) and RT-PCR. The changes in the biological characteristics of the transfected MSCs were also observed.

RESULTS

Stable overexpression of bFGF protein was detected in the transfected BMSCs, which showed differentiation towards chondrocyte lineage.

CONCLUSION

Stable expression of bFGF gene in transfected BMSCs can induce cell differentiation into chondrocyte lineage.

摘要

目的

探讨将碱性成纤维细胞生长因子(bFGF)基因转入兔骨髓基质细胞(BMSCs)的方法及可能性。

方法

构建携带bFGF cDNA的真核表达载体,通过脂质体介导转染兔BMSCs。采用形态学观察、免疫组织化学、酶联免疫吸附测定(ELISA)及逆转录-聚合酶链反应(RT-PCR)检测转染后BMSCs中bFGF基因的转录和表达情况。同时观察转染后间充质干细胞生物学特性的变化。

结果

在转染的BMSCs中检测到bFGF蛋白的稳定过表达,且这些细胞向软骨细胞系分化。

结论

bFGF基因在转染的BMSCs中的稳定表达可诱导细胞向软骨细胞系分化。

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