CAJ | 학술논문

目的探讨Sox9基因转染对骨髓间充质干细胞(MSCs)成软骨分化的影响.方法采用密度梯度离心和贴壁培养法分离兔骨髓MSCs,体外培养扩增.阳离子脂质体介导重组真核表达Sox9质粒转染骨髓MSCs,通过反转录-聚合酶链反应(RT-PCR)和蛋白印迹实验(Westem blot)的方法检测基因产物的表达,荧光显微镜和流式细胞术测定细胞转染效率;四甲基噻唑蓝(MTT)法测定基因转染对细胞增殖能力的影响,流式细胞术对转基因细胞进行细胞周期分析;通过Westem blot,RT-PCR和免疫组织化学染色的方法检测Sox9基因转染对MSCs成软骨分化的影响.结果脂质体介导Sox9基因可以成功地转染兔骨髓MSCs;Sox9基因转染对细胞增殖能力无明显影响;基因转染后的细胞表达软骨分化特异性分子Ⅱ型胶原,Sox9,aggrecan等明显增加.结论 Sox9基因转染骨髓MSCs可以获得稳定表达,基因表达产物可以促进骨髓MSCs向软骨方向的分化.
목적 탐토Sox9기인전염대골수간충질간세포(MSCs)성연골분화적영향.방법 채용밀도제도리심화첩벽배양법분리토골수MSCs,체외배양확증.양리자지질체개도중조진핵표체Sox9질립전염골수MSCs,통과반전록-취합매련반응(RT-PCR)화단백인적실험(Westem blot)적방법검측기인산물적표체,형광현미경화류식세포술측정세포전염효솔;사갑기새서람(MTT)법측정기인전염대세포증식능력적영향,류식세포술대전기인세포진행세포주기분석;통과Westem blot,RT-PCR화면역조직화학염색적방법검측Sox9기인전염대MSCs성연골분화적영향.결과 지질체개도Sox9기인가이성공지전염토골수MSCs;Sox9기인전염대세포증식능력무명현영향;기인전염후적세포표체연골분화특이성분자Ⅱ형효원,Sox9,aggrecan등명현증가.결론 Sox9기인전염골수MSCs가이획득은정표체,기인표체산물가이촉진골수MSCs향연골방향적분화.
Objective To observe the effect of Sox9 gene overexpressian on chondrogenesis of esenchymal stem ceils (MSCs) in vitro. Methods Rabbit MSCs were obtained and purified by gradient centrifuge and adhesion culture in vitro. MSCs were transfected by recombinant Sox9 plasmid. RT-PCR and Western blot analysis were used for transfection confirmation. Chondrogenic molecules such as type I1 collagen,aggrecan and Sox9 were detected by immunohistology, RT-PCR and western blot. Cell proliferation was tested by MTT assay. Results MSCs were successfully transfected with Sox9 gene and verified by RT-PCR and Western blot analysis. The overexpression of Sox9 had no effect on the proliferation of MSCs. Transfeeted cells expressed more chondrogenic markers than the control groups. Conclusion Sox9 gene overexpression can enhance chondrogenic differentiation of MSCs.