SOX-9真核表达质粒转染兔骨髓基质细胞

Experimental study on the bone marrow stromal cells of rabbits transfected by SOX-9 gene

目的探讨SOX(sry-type high-mobility-group box)-9基因转染兔骨髓基质细胞的方法及可行性。方法构建SOX-9基因真核表达质粒,用脂质体法介导转染兔骨髓基质细胞,通过形态学观察、免疫组织化学、酶联免疫吸附法及反转录-聚合酶链反应检测SOX-9基因转染兔骨髓基质细胞的成功性,以及转染后骨髓基质干细胞的生物学特性。结果免疫组织化学检测实验组细胞内有稳定的SOX-9表达,在2周的表达量高于6d的表达量。RT-PCR结果表明只有实验组有SOX-9的基因表达。酶联免疫吸附法检测结果显示各时间点SOX-9的基因表达实验组均明显高于其余2组(P<0.01),实验组表达量48h时达到最高。结论SOX-9基因可以转染兔骨髓基质干细胞,并能诱导骨髓基质干细胞向成软骨方向分化。

Objective To investigate the method and feasibility of transfection on SOX-9 gene into marrow stroma cell （BMSCs） of rabbits. Methods The eukaryotic expression vectors that were constructed by SOX-9 cDNA were transfected into BMSCs of rabbits by liposome induction. The transcription and expression of SOX-9 gene in BMSCs were detected by morphology observation mean, immunohistochemistry,enzyme linked immunosorbent assay and reverse transcription-polymerasechain reaction （RT-PCR） means. The changes of biologic characterisctics of transfected BMSCs were also observed. Results The stable expression of SOX-9 proteins was detected in the experimental group by immunohistochemistry, and it was higher in 2 weeks than in 6 days. The expression of SOX-9 gene were observed only in experimental group by RT-PCR means. The expression of SOX-9 protein were detected in three group by ELISA means, and it was remarkably higher in the experimental group than other group （P 〈 0.01 ）, and it was the highest at 48h in the experimental group. Conclusion The BMSCs of rabbits can be transfected and are induced to differentiate towards cartilage by SOX-9 gene.