The Comparative Study of Different Methods for Gene Transfer to Cochlear Hair Cells
10.3969/j.issn.1006-7299.2015.02.012
- VernacularTitle:不同基因转染方法对小鼠内耳毛细胞转染效率的比较
- Author:
Yan CHEN
;
Chuijin LAI
;
Wen LI
;
Huawei LI
- Publication Type:Journal Article
- Keywords:
Inner ear;
Cochlear hair cells;
Gene transfer;
Electroporation;
Adenovector;
Lentivirus vector
- From:
Journal of Audiology and Speech Pathology
2015;(2):166-169
- CountryChina
- Language:Chinese
-
Abstract:
Objective To obtain an easy and high efficient method for gene transfer to cochlear hair cells ,by comparing three mediating green fluorescent protein (GFP) methods (electroporation ,adenovector and lentivirus vector) .Methods Cochlear sensory epithelium was dissected from anaesthetized P2 mice .Sensory epithelia were transferred onto poly -L -lysine treated cover slides and cultured overnight .Gene transfer was performed by elec‐troporation in medium containing pGPHI/GFP/Neo plasmid or by incubation with diluted recombined adenovirus/lentivirus vector .After 48 hours ,green fluorescence was checked under fluorescence microscope .To confirm the ef‐ficiency of exogenous gene transfer ,real-time PCR was performed using specific primers .Results The transfec‐tion efficiencies of electroporation and lentivirus vector mediated gene transfer were very low .Both immunofluores‐cence and real - time PCR results showed that the transfection efficiency of adenovirus mediated GFP and Bmi1 transfer were relatively higher .The proportion of GFP positive cells in outer hair cells and inner hair cells of middle turn were 90 .0% ± 4 .1% and 5% ± 0 .4% ,respectively .Conclusion Adenovector is more efficient for exogenous gene transfer to cochlear hair cells ,thus adenovector is a good carrier for gene transfer to cochlear hair cells .
