本文關(guān)鍵詞： CD47 骨髓間充質(zhì)干細胞 重組質(zhì)粒 轉(zhuǎn)染 表達　出處：《重慶醫(yī)科大學》2014年碩士論文　論文類型：學位論文

【摘要】：目的：構(gòu)建CD47重組質(zhì)粒并將其傳染至大鼠骨髓間充質(zhì)干細胞(bone mesenchymal stem cells, BMSCs)，建立穩(wěn)定過表達CD47的BMSCs，為研究干細胞治療中逃避免疫監(jiān)視的有關(guān)機制提供實驗模型。 方法：構(gòu)建過表達CD47基因的質(zhì)粒載體，轉(zhuǎn)染至分離培養(yǎng)的大鼠BMSCs，，RT-PCR、Western blot分別檢測干細胞CD47mRNA、蛋白水平的表達，并完成功能檢測。 結(jié)果：成功構(gòu)建了pYr-ads-4-ratCD47質(zhì)粒，經(jīng)酶切、測序結(jié)果表明pYr-ads-4-ratCD47重組質(zhì)粒構(gòu)建成功；RT-PCR、Western blot顯示實驗組轉(zhuǎn)染至大鼠BMSCs后CD47的mRNA和蛋白水平表達明顯增高（p0.05）；功能檢測提示過表達CD47分子的BMSCs成功逃避巨噬細胞吞噬。 結(jié)論：經(jīng)質(zhì)粒介導可成功將大鼠CD47目的基因片段轉(zhuǎn)染至大鼠BMSCs，并能夠在體外有效的過表達。
[Abstract]:Objective : To construct CD47 recombinant plasmid and transfer it to bone marrow mesenchymal stem cells ( MSCs ) in rat bone marrow mesenchymal stem cells ( MSCs ) , and to establish bone marrow cells stably expressing CD47 to provide experimental model for the study of immune surveillance in stem cell therapy . Methods : The expression of CD47mRNA and protein in stem cells was detected by RT - PCR and Western blot , and the function of CD47 mRNA was detected by RT - PCR and Western blot . Results : The expression of CD47 was successfully constructed by RT - PCR and Western blot , and the expression of CD47 mRNA and protein was significantly increased ( p < 0.05 ) . Conclusion : The gene fragment of rat CD47 can be successfully transfected into rat bone marrow cells via plasmid - mediated gene transfer , and can be effectively expressed in vitro .

【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R392

【參考文獻】

相關(guān)期刊論文 前1條

1 王穎超;馮磊;殷楚云;馬麗娜;魏永緯;王春美;盛光耀;;抗CD47抗體聯(lián)合阿糖胞苷靶向治療NOD/SCID小鼠單核細胞白血病的研究[J];中國當代兒科雜志;2013年07期

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