發(fā)布時(shí)間：2018-01-30 20:58

  本文關(guān)鍵詞： CD47 骨髓間充質(zhì)干細(xì)胞 重組質(zhì)粒 轉(zhuǎn)染 表達(dá)　出處：《重慶醫(yī)科大學(xué)》2014年碩士論文　論文類(lèi)型：學(xué)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(xì)胞(bone mesenchymal stem cells, BMSCs)，建立穩(wěn)定過(guò)表達(dá)CD47的BMSCs，為研究干細(xì)胞治療中逃避免疫監(jiān)視的有關(guān)機(jī)制提供實(shí)驗(yàn)?zāi)Ｐ汀?方法：構(gòu)建過(guò)表達(dá)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(cè)干細(xì)胞CD47mRNA、蛋白水平的表達(dá)，并完成功能檢測(cè)。 結(jié)果：成功構(gòu)建了pYr-ads-4-ratCD47質(zhì)粒，經(jīng)酶切、測(cè)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(shí)驗(yàn)組轉(zhuǎn)染至大鼠BMSCs后CD47的mRNA和蛋白水平表達(dá)明顯增高（p0.05）；功能檢測(cè)提示過(guò)表達(dá)CD47分子的BMSCs成功逃避巨噬細(xì)胞吞噬。 結(jié)論：經(jīng)質(zhì)粒介導(dǎo)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(guò)表達(dá)。
[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(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R392

【參考文獻(xiàn)】

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(xì)胞白血病的研究[J];中國(guó)當(dāng)代兒科雜志;2013年07期

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