Method for differentiating synovium mesenchymal stem cells (SMSCs) to chondrocytes through in-vitro adenovirus mediated BMP-2/7 gene coexpression induction

体外腺病毒介导bmp-2/7基因共表达诱导滑膜间充质干细胞向软骨细胞分化的方法

Abstract

The invention relates to a method for differentiating SMSCs to chondrocytes through in-vitro adenovirus mediated BMP-2/7 gene coexpression induction. The method comprises the following steps: separating and culturing SMSCs; constructing a pAdTrack-CMV-BMP2-IRES-BMP7 double gene recombinant vector; transferring the fifth-generation SMSCs through utilizing the pAdTrack-CMV-BMP2-IRES-BMP7 double gene recombinant vector; amplifying the recombinant viruses obtained after the transfection; adding the recombinant viruses to a chondroblast induction culture solution for induction; and culturing for above 10 days to obtain fibrochondrocytes. The method which adopts the adenovirus mediated BMP-2/7 gene coexpression vector to transfer SMSCs has the advantages of high transgenic efficiency, instant expression and high safety, and allows the SMSCs to be differentiated to the chondrocytes through the in-vitro induction.
本发明一种体外腺病毒介导BMP-2/7基因共表达诱导滑膜间充质干细胞向软骨细胞分化的方法,包括一个将滑膜间充质干细胞分离和培养的步骤;一个构建pAdTrack-CMV-BMP2-IRES-BMP7双基因重组载体的步骤,一个利用pAdTrack-CMV-BMP2-IRES-BMP7双基因重组载体转染第五代滑膜间充质干细胞的步骤,一个将转染后获得的重组病毒扩增的步骤;一个将上述的重组病毒加入成软骨细胞诱导培养液中诱导的步骤,培养10天以上即可获得纤维软骨样细胞。本发明采用腺病毒介导BMP-2/7共表达载体转染SMSCs,转基因效率高,瞬间表达,安全性高,在体外诱导SMSCs向纤维软骨细胞分化。

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