|本期目录/Table of Contents|

[1]高 鹏,张 春,赵 巍,等.重组腺病毒载体对大鼠胶质瘤细胞转染效率的影响[J].宁夏医科大学学报,2013,(12):1322-1325,1332,1302.
 GAO Peng,ZHANG Chun,ZHAO Wei,et al.Infection Efficiency of Rat C6 Cells Infected with Recombinant Adenovirus Vectors[J].Ningxia Medical University,2013,(12):1322-1325,1332,1302.
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《宁夏医科大学学报》[ISSN:1005-8486/CN:64-1029/R]

卷:
期数:
2013年12期
页码:
1322-1325,1332,1302
栏目:
论著
出版日期:
2013-12-20

文章信息/Info

Title:
Infection Efficiency of Rat C6 Cells Infected with Recombinant Adenovirus Vectors
作者:
高 鹏12 张 春3 赵 巍4 沈 冰2
1.宁夏医科大学,银川 750004; 2.宁夏医科大学总医院神经外科,银川 750004; 3.宁夏医科大学颅脑重点实验室,银川 750004; 4.宁夏医科大学医学科学技术研究中心,银川 750004
Author(s):
GAO Peng12 ZHANG Chun3 ZHAO Wei4 SHEN Bing2
1.Ningxia Medical University, Yinchuan 750004; 2.Department of Neurosurgery, the General Hospital of Ningxia Medical University, Yinchuan 750004; 3.Ningxia Key Laboratory of Cerebrocranial Diseases, Yinchuan 750004; 4. Medical Sci-Tech Research Center of Ningxia Medical University, Yinchuan 750004
关键词:
腺病毒转染大鼠胶质瘤细胞转染复数
Keywords:
adenovirus infection rat glioma cells infection efficiency
分类号:
R739.41
DOI:
-
文献标志码:
A
摘要:
目的研究转染复数(MOI)和转染时间对重组腺病毒载体转染大鼠胶质瘤细胞转染效率的影响。方法体外培养大鼠胶质瘤细胞(C6细胞),用携带有rHSG基因的重组腺病毒载体(Adv-rHSG-GFP)转染C6细胞。根据MOI将细胞分为:A组MOI=0、B组MOI=10、C组MOI=50、D组MOI=80、E组MOI=100,转染Adv-rHSG-GFP24h后分别收集每组细胞,采用WesternBlot和免疫细胞化学法检测rHSG蛋白的表达,并通过流式细胞仪和倒置荧光显微镜观察GFP阳性表达来评价重组腺病毒载体的转染效率。用MOI为100的Adv-rHSG-GFP转染C6细胞不同时间后(12、24、36和48h),同法收集细胞并检测rHSG蛋白的表达及评价重组腺病毒载体的转染效率。结果Adv-rHSG-GFP转染C6细胞24h后,rHSG蛋白的相对表达量和绿色荧光蛋白表达随MOI值的增高而增多;用MOI为100的Adv-rHSG-GFP转染C6细胞后,12h即可观察到绿色荧光蛋白的表达,且在24h内随时间的变化而增高,在24~36h绿色荧光蛋白表达无变化,48h有所降低(P<0.01);rHSG蛋白相对表达量在12~48h内随时间的变化而增高,于48h达到最大值(P<0.01)。结论重组腺病毒载体能够有效转染C6细胞,且在MOI值为100,24h时有最高转染效率。
Abstract:
Objective To investigate the effect of multiplicity of infection(MOI)and infection time on infection efficiency of rat C6 cells infected with recombinant adenovirus vectors. Methods The rat glioma cells(C6 cells)were cultured in vitro and infected with recombinant adenovirus vectors containing rHSG(Adv-rHSG-GFP). Based on the MOI values, the cells were divided into group A(MOI=0), group B(MOI=10), group C(MOI=50), group D(MOI=80)and group E(MOI=100). Cells of different groups were collected and tested after infection for 24h. Western blot and immunohistochemical methods were applied for determining the expression of rHSG protein; flow cytometry(FCM)and inverted fluorescence microscope were used for observing GFP positive expression to assess the infection efficiency of adenovirus vectors. Then C6 cells were infected with Adv-rHSG-GFP(MOI=100)for different time periods(12h, 24h, 36h, 48h)and collected. rHSG protein expression was detected and infection efficiency was evaluated in the same manner. Results It was observed that in C6 cells infected with Adv-rHSG-GFP for 24h, the protein expression of rHSG and green fluorescent protein were increasing with MOI. After infection with Adv-rHSG-GFP(MOI=100), green fluorescent protein could be observed after 12h and increased as time went on within 24h while no change from 24~36h and decreased after 48h(P<0.01). rHSG protein expression level raised within the range of 12~48h and maximized at 48h(P<0.01). Conclusion The recombinant aden ovirus vectors can infect C6 cells effectively, and the highest infection efficiency can be obtained at MOI=100 and 24h.

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备注/Memo

备注/Memo:
收稿日期:2013-09-05 基金项目:宁夏教育厅基金资助(NGY2011054) 作者简介:高鹏(1986-),男,在读硕士研究生,从事神经肿瘤研究。 通信作者:沈冰,医学博士,副教授,硕士研究生导师。 E-mail:shenbing315@sina.com
更新日期/Last Update: 2013-12-20