[1]吕颖慧,王启钊,刁勇,等.脂质体转染人肝星状细胞和肝细胞条件优化[J].华侨大学学报(自然科学版),2010,31(1):49-52.[doi:10.11830/ISSN.1000-5013.2010.01.0049]
　Lü Ying-hui,WANG Qi-zhao,DIAO Yong,et al.Optimization of the Transfection Efficiency of Human Hepatic Stellate Cell and Hepatocyte Using Liposome[J].Journal of Huaqiao University(Natural Science),2010,31(1):49-52.[doi:10.11830/ISSN.1000-5013.2010.01.0049]
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脂质体转染人肝星状细胞和肝细胞条件优化()
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《华侨大学学报（自然科学版）》[ISSN:1000-5013/CN:35-1079/N]

卷:
第31卷
期数:
2010年第1期
页码:
49-52
栏目:
出版日期:
2010-01-20

文章信息/Info

Title:
Optimization of the Transfection Efficiency of Human Hepatic Stellate Cell and Hepatocyte Using Liposome
文章编号:
1000-5013(2010)01-0049-04
作者:
吕颖慧; 王启钊; 刁勇; 许瑞安
华侨大学分子药物学研究所
Author(s):
Lü Ying-hui;  WANG Qi-zhao;  DIAO Yong;  XU Rui-an
Institute of Molecular Medicine, Huaqiao University, Quanzhou 362021, China
关键词:
脂质体;  肝星状细胞;  肝细胞;  转染效率
Keywords:
liposome;  hepatic stellate cells;  hepatocyte;  transfection efficiency
分类号:
Q78
DOI:
10.11830/ISSN.1000-5013.2010.01.0049
文献标志码:
A
摘要:
利用Lipofectamine 2000(Lipo)介导LacZ及EGFP基因转染人肝星状细胞系LX-2和人肝细胞系Chang Liver,探讨Lipo用量、质粒用量、转染时间、细胞初始接种密度及不同启动子驱动对转染效率的影响.结果显示,两种细胞均能得到高效转染,Chang Liver细胞转染效率(>80%)高于LX-2细胞(>60%).Lipo用量为每孔2μL,转染后6 h换液,LX-2细胞应用质粒DNA为每孔0.45μg,Chang Liver细胞应用质粒DNA为每孔0.30~0.45μg,此时转染效率最高.巨细胞病毒(CMV)启动子驱动LacZ转染效率与CMV早期增强子/鸡β肌动蛋白(CAG)启动子驱动增强型绿色荧光蛋白(EGFP)转染效率无显著差异; 细胞初始接种密度对转染效率无显著影响.
Abstract:
PAAV-CMV-LacZ plasmid and pAAV-CAG-EGFP plasmid were transfected into a kind of human hepatic stellate cell line-LX-2 and a kind of human hepatocyte line-Chang Liver using Lipofectamine 2000,respectively.The impacts of liposome and plasmid amount,transfection time course,initial plated density and different promoter used on transfection efficiency were discussed.It indicated that both kinds of cell lines were highly transfectable,and the transfection efficiency of Chang Liver cells(>80%) was higher than LX-2 cells(>60%).The transfection efficiency turned out to be the highest while using 2 μL·well-1 of liposome,0.45 μg·well-1 of plasmid DNA for LX-2 cells and 0.30~0.45 μg·well-1 of plasmid DNA for Chang Liver cells after 6 hours of transfection.There was no significantly difference in transfection efficiency whether the gene was driven by CMV or CAG promoter.The initial plated density seemed to have no obvious influence on the transfection efficiency,either.

参考文献/References:

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备注/Memo

备注/Memo:
厦门市科技计划项目(3502Z20053046); 泉州市科技计划项目(2007Z41); 华侨大学高层次人才科研启动项目(09BS516)
更新日期/Last Update: 2014-03-23