SOX9基因修饰骨髓间充质干细胞并诱导其向髓核样细胞分化的实验研究
发布时间:2018-06-10 19:05
本文选题:间质干细胞 + 慢病毒感染 ; 参考:《重庆医学》2013年01期
【摘要】:目的探讨SOX9慢病毒载体感染兔骨髓间充质干细胞(MSC)并诱导其向髓核样细胞的分化。方法构建SOX9基因慢病毒载体并感染MSC,将MSC分为未转导组、空转导组及SOX9转导组。采用流式细胞仪检测MSC的感染率及凋亡率,MTT检测MSC的增殖情况,RT-PCR、Western blot及免疫荧光染色检测MSC中SOX9、Ⅱ型胶原及Aggrecan在mRNA及蛋白水平的表达情况。结果成功构建SOX9基因慢病毒载体并感染MSC,感染率为100%。慢病毒载体感染对3组MSC的增殖及凋亡无明显影响;感染2周后,SOX9转导组MSC中SOX9及Ⅱ型胶原在mRNA及蛋白水平的表达均明显高于未转导组与空转导组(P0.01),Aggrecan mRNA及蛋白仅在SOX9转导组MSC中表达;SOX9转导组MSC中Ⅱ型胶原及Aggrecan荧光染色呈阳性,而空转导组及未转导组MSC均呈阴性。结论 SOX9慢病毒载体感染的MSC可被诱导向髓核细胞分化。
[Abstract]:Objective to investigate the differentiation of bone marrow mesenchymal stem cells (MSCs) infected with SOX9 lentivirus vector. Methods SOX9 lentivirus vector was constructed and infected with MSC.MSC was divided into three groups: untransduced group, empty transduction group and SOX9 transduction group. Flow cytometry was used to detect the infection rate and apoptosis rate of MSCs. The proliferation of MSCs was detected by MTT. The expression of SOX9, type 鈪,
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