SOX9基因修飾骨髓間充質(zhì)干細胞并誘導其向髓核樣細胞分化的實驗研究
發(fā)布時間:2018-06-10 19:05
本文選題:間質(zhì)干細胞 + 慢病毒感染 ; 參考:《重慶醫(yī)學》2013年01期
【摘要】:目的探討SOX9慢病毒載體感染兔骨髓間充質(zhì)干細胞(MSC)并誘導其向髓核樣細胞的分化。方法構(gòu)建SOX9基因慢病毒載體并感染MSC,將MSC分為未轉(zhuǎn)導組、空轉(zhuǎn)導組及SOX9轉(zhuǎn)導組。采用流式細胞儀檢測MSC的感染率及凋亡率,MTT檢測MSC的增殖情況,RT-PCR、Western blot及免疫熒光染色檢測MSC中SOX9、Ⅱ型膠原及Aggrecan在mRNA及蛋白水平的表達情況。結(jié)果成功構(gòu)建SOX9基因慢病毒載體并感染MSC,感染率為100%。慢病毒載體感染對3組MSC的增殖及凋亡無明顯影響;感染2周后,SOX9轉(zhuǎn)導組MSC中SOX9及Ⅱ型膠原在mRNA及蛋白水平的表達均明顯高于未轉(zhuǎn)導組與空轉(zhuǎn)導組(P0.01),Aggrecan mRNA及蛋白僅在SOX9轉(zhuǎn)導組MSC中表達;SOX9轉(zhuǎn)導組MSC中Ⅱ型膠原及Aggrecan熒光染色呈陽性,而空轉(zhuǎn)導組及未轉(zhuǎn)導組MSC均呈陰性。結(jié)論 SOX9慢病毒載體感染的MSC可被誘導向髓核細胞分化。
[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,
本文編號:2004279
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