比较携带IDO腺病毒转染肝细胞的三种方法

发布时间：2018-06-12 07:57

本文选题：IDO + 腺病毒　；参考：《福建医科大学》2012年硕士论文

【摘要】：目的：研究三种转染途径对携带吲哚胺2，3－二氧化酶（indoleaminc2,3-dioxygcnasc,IDO)腺病毒转染小鼠肝细胞的效率，肝细胞活性及IDO基因的表达。方法：用相同量携带IDO基因的重组腺病毒通过三种方式转染BABL/c小鼠的肝脏，采用胶原酶在体灌注和离体消化的方法分离小鼠的肝细胞，用台盼蓝拒染法来检测肝细胞活性，使用RT-PCR和Western Blot法分别检测转染后的肝细胞内IDO基因及其蛋白的表达。结果：门静脉灌注加离心法成功分离并纯化肝细胞，肝细胞数量1.62×108／只。腺病毒成功转染肝细胞，三组平均转染率分别为：体外培养组27.298%，肝细胞活性为79.076%；门静脉组24.676%，肝细胞活性为94.768%；外周静脉组21.01%，肝细胞活性为90.812%。RT-PCR、WestBlot可以检测到IDO基因在肝细胞内成功表达。结论：经门静脉注射组可获得高转染率，，高活性的肝细胞。
[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.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R373

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