本文關(guān)鍵詞：三種攜帶GFP慢病毒轉(zhuǎn)染兔骨髓間充質(zhì)干細(xì)胞的效果　出處：《中國(guó)組織工程研究》2017年29期 　論文類(lèi)型：期刊論文

  更多相關(guān)文章： 慢病毒屬 骨髓 間質(zhì)干細(xì)胞 綠色熒光蛋白質(zhì)類(lèi) 組織工程 干細(xì)胞 骨髓干細(xì)胞 綠色熒光蛋白 間充質(zhì)干細(xì)胞 骨髓間充質(zhì)干細(xì)胞 慢病毒 轉(zhuǎn)染 轉(zhuǎn)染率 生長(zhǎng)曲線(xiàn) 國(guó)家自然科學(xué)基金

【摘要】：背景:研究不同慢病毒對(duì)同一骨髓來(lái)源間充質(zhì)干細(xì)胞的轉(zhuǎn)染效果,篩選合適的細(xì)胞轉(zhuǎn)染工具。目的:探究3種慢病毒對(duì)兔骨髓間充質(zhì)干細(xì)胞轉(zhuǎn)染效果的差異。方法:選擇2周齡日本大耳白兔,髂骨穿刺抽取骨髓液,采用密度梯度離心法提取骨髓間充質(zhì)干細(xì)胞,流式細(xì)胞儀檢測(cè)細(xì)胞表面標(biāo)志物表達(dá)。分別用p Tomo-GFP慢病毒、Ad-GFP慢病毒、lenti-GFP慢病毒轉(zhuǎn)染骨髓間充質(zhì)干細(xì)胞,流式細(xì)胞儀檢測(cè)3種慢病毒的轉(zhuǎn)染率。CCK-8法檢測(cè)轉(zhuǎn)染前后細(xì)胞生長(zhǎng)活力變化。結(jié)果與結(jié)論:①培養(yǎng)出的細(xì)胞CD44陽(yáng)性率為75.64%,CD34、CD45陽(yáng)性率分別為6.14%、9.48%;②pTomo-GFP慢病毒轉(zhuǎn)染后GFP表達(dá)率為2.64%、Ad-GFP慢病毒轉(zhuǎn)染后GFP表達(dá)率為16.72%、lenti-GFP慢病毒轉(zhuǎn)染后GFP表達(dá)率為45.24%;③各慢病毒對(duì)細(xì)胞生長(zhǎng)存在一定影響。p Tomo-GFP、Ad-GFP在第1-8天對(duì)細(xì)胞生長(zhǎng)潛伏期和對(duì)數(shù)生長(zhǎng)期存在影響,可能對(duì)前期細(xì)胞活性造成一定干擾。短期內(nèi)lenti-GFP對(duì)細(xì)胞生長(zhǎng)影響不大,但第12天時(shí)出現(xiàn)了細(xì)胞生長(zhǎng)情況變化,考慮對(duì)細(xì)胞遠(yuǎn)期活力存在影響;④選用lenti-GFP慢病毒轉(zhuǎn)染兔骨髓間充質(zhì)干細(xì)胞能夠在保證細(xì)胞正常生長(zhǎng)的情況下提供最大的轉(zhuǎn)染效率,適用于兔骨髓間充質(zhì)干細(xì)胞的示蹤及基因轉(zhuǎn)染等相關(guān)研究。
[Abstract]:Background: to study the transfection effect of different lentiviruses on mesenchymal stem cells from the same bone marrow source. Objective: to investigate the transfection effects of three lentiviruses on rabbit bone marrow mesenchymal stem cells (BMSCs). Methods: two week-old Japanese large ear white rabbits were selected and bone marrow fluid was extracted by iliac puncture. Bone marrow mesenchymal stem cells (BMSCs) were extracted by density gradient centrifugation, and the expression of cell surface markers was detected by flow cytometry. Lenti-GFP lentivirus was transfected into bone marrow mesenchymal stem cells. Flow cytometry was used to detect the transfection rate of three lentiviruses. CCK-8 method was used to detect the changes of cell growth activity before and after transfection. Results and conclusion the CD44 positive rate of the cells cultured at 1: 1 was 75.64%. The positive rate of CD34 + CD45 was 6.14% and 9.48%, respectively. 2the GFP expression rate of pTomo-GFP lentivirus was 2.64% and the GFP expression rate of Ad-GFP lentivirus transfection was 16.72%. The expression rate of GFP was 45.24% after lenti-GFP lentivirus transfection. (3) Lentivirus had a certain effect on cell growth. P Tomo-GFP Ad-GFP had an effect on cell growth latency and logarithmic growth period on the 1st to 8th day. Lenti-GFP had little effect on cell growth in the short term, but the change of cell growth appeared on the 12th day. (4) transfection of rabbit bone marrow mesenchymal stem cells with lenti-GFP lentivirus could provide the maximum transfection efficiency under the condition of ensuring the normal growth of the cells. It is suitable for tracing and gene transfection of rabbit bone marrow mesenchymal stem cells.
【作者單位】： 昆明醫(yī)科大學(xué)第一附屬醫(yī)院運(yùn)動(dòng)醫(yī)學(xué)科;
【基金】：國(guó)家自然科學(xué)基金(81460340) 云南省骨關(guān)節(jié)疾病診療省創(chuàng)新團(tuán)隊(duì)項(xiàng)目(2014HC1018) 醫(yī)療衛(wèi)生內(nèi)設(shè)機(jī)構(gòu)研究項(xiàng)目(2014NS161)~~
【分類(lèi)號(hào)】：R329.2
【正文快照】： 文章快速閱讀:文題釋義:綠色熒光蛋白:發(fā)現(xiàn)于維多利亞水母,是由238個(gè)氨基酸組成,經(jīng)紫外光激發(fā)后可發(fā)出峰值為509 nm的綠色熒光,廣泛用于基因工程中的基因表達(dá)檢測(cè)、細(xì)胞示蹤等。經(jīng)過(guò)改良,研制出了增強(qiáng)型綠色熒光蛋白及其他顏色的熒光蛋白變體。慢病毒:屬反轉(zhuǎn)錄病毒,具有長(zhǎng)期潛

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