本文選題：IDO + 腺病毒　； 參考：《福建醫(yī)科大學(xué)》2012年碩士論文

【摘要】：目的：研究三種轉(zhuǎn)染途徑對(duì)攜帶吲哚胺2，3－二氧化酶（indoleaminc2,3-dioxygcnasc,IDO)腺病毒轉(zhuǎn)染小鼠肝細(xì)胞的效率，肝細(xì)胞活性及IDO基因的表達(dá)。 方法：用相同量攜帶IDO基因的重組腺病毒通過三種方式轉(zhuǎn)染BABL/c小鼠的肝臟，采用膠原酶在體灌注和離體消化的方法分離小鼠的肝細(xì)胞，用臺(tái)盼藍(lán)拒染法來檢測肝細(xì)胞活性，使用RT-PCR和Western Blot法分別檢測轉(zhuǎn)染后的肝細(xì)胞內(nèi)IDO基因及其蛋白的表達(dá)。 結(jié)果：門靜脈灌注加離心法成功分離并純化肝細(xì)胞，肝細(xì)胞數(shù)量1.62×108／只。腺病毒成功轉(zhuǎn)染肝細(xì)胞，三組平均轉(zhuǎn)染率分別為：體外培養(yǎng)組27.298%，肝細(xì)胞活性為79.076%；門靜脈組24.676%，肝細(xì)胞活性為94.768%；外周靜脈組21.01%，肝細(xì)胞活性為90.812%。RT-PCR、WestBlot可以檢測到IDO基因在肝細(xì)胞內(nèi)成功表達(dá)。 結(jié)論：經(jīng)門靜脈注射組可獲得高轉(zhuǎn)染率，，高活性的肝細(xì)胞。
[Abstract]:Objective: to study the efficiency of three transfection pathways on the transfection of mouse hepatocytes by adenovirus carrying indoleaminc2n 3-dioxygcnascan IDO. Methods: the recombinant adenovirus carrying IDO gene was transfected into the liver of Babl / c mice in three ways, and the hepatocytes were isolated by collagenase perfusion and in vitro digestion. The activity of hepatocytes was detected by trypan blue exclusion method, and the expression of IDO gene and its protein in transfected hepatocytes was detected by RT-PCR and Western Blot. Results: hepatocytes were isolated and purified by portal vein perfusion and centrifugation, the number of hepatocytes was 1.62 脳 108 / mouse. Adenovirus was successfully transfected into hepatocytes. The average transfection rates of the three groups were 27.298 in vitro, 79.076 in liver cell activity, 94.768 in portal vein group and 21.01 in peripheral vein group. High transfection efficiency was obtained in portal vein injection group. Highly active hepatocytes.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R373

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相關(guān)期刊論文 前3條

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本文編號(hào)：2008959