Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (19): 3507-3512.doi: 10.3969/j.issn.1673-8225.2010.19.018

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Identification of human bone marrow mesenchymal stem cells transfected with basic fibroblast growth factor gene in vitro

Zheng You-hua, Zhang Zhi-guang, Su Kai, Kuang Shi-jun   

  1. Department of Oral and Maxillofacial Surgery, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou   510055, Guangdong Province, China
  • Online:2010-05-07 Published:2010-05-07
  • About author:Zheng You-hua, Doctor, Associate professor, Associate chief physician, Department of Oral and Maxillofacial Surgery, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou 510055, Guangdong Province, China zhengyhgz@tom.com
  • Supported by:

    the Social Development Program of Guangdong Provincial Science and Technology Department, No. 2008B030301312*

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Abstract:

BACKGROUND: Directional differentiation of bone marrow mesenchymal stem cells (BMSCs) requires regulation of appropriate growth factors. Increasing attention has been paid to transfect cells with cytokine gene for promoting BMSCs to proliferate and differentiate and to accelerate repair process of bone defects.
OBJECTIVE: To investigate the biological characteristics of BMSCs as seed cells of tissue engineering after transfected with basic fibroblast growth factor (bFGF) gene in vitro.
METHODS: The phagemid expression vector with human pcDNA3.1-bFGF was amplified in E. coli. The plasmid was extracted, purified by EndoFree Plasmid Maxi Kit. The plasmid of pcDNA3.1-bFGF was analyzed and identified by enzymes digestion and sequencing analysis. pcDNA3.1-bFGF gene was transfected into P3 BMSCs using Lipofectamine 2000. Positive clones of BMSCs transfected bFGF gene were selected with G418. The expression of bFGF mRNA and its productions in the transfected BMSCs were detected by real time PCR, immunohistochemistry, immunofluorescence and Western-blot. The proliferative cycle of transfected BMSCs were examined by flow cytometry analysis.
RESULTS AND CONCLUSION: BMSCs expressing bFGF gene were obtained by transfection with pcDNA3.1-bFGF gene via Lipofectamine. The cell lineage was confirmed that the expressed position of bFGF was mainly in cytoplasm after transfection with bFGF gene. Higher proportion of cells in proliferation cycle was shown in transfected BMSCs compared with non-transfected BMSCs (P < 0.05). Results show that pcDNA3.1-bFGF can be transfected into BMSCs cultured in vitro via Lipofectamine. Genetic modification of BMSCs with bFGF may promote the proliferation of BMSCs.

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