TITLE

人永生化表皮细胞HaCaT电转染条件的优化

AUTHOR(S)
徐 珊; 杜 冲; 马艳民; 谢宏俊; 高杨; 石 琦; 王新阳; 贺大林; 郭 鹏
PUB. DATE
July 2014
SOURCE
Journal of Xi'an Jiaotong University (Medical Sciences);Jul2014, Vol. 35 Issue 5, p551
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
Objective To investigate the method and optimum conditions of electrotransfection and the major influential factors of electroransfection efficiency and the survival rate of human immortalized epidermal keratinization cells. Methods The pGPU6/GFP/nev plasmids were electrotransfected into HaCaT by electroporation apparatus with different electric voltages (180,360,540 and 720 V),time of pulse (40,60,80,100 and 120μs),electronic shock times (1,2,3,4 and 5),and incubation temperature (room temperature and 4 °C). The number of green fluorescence positive cells and the total cell number were counted respectively under fluorescent microscope and visible light microscope after 24h,the electrotransfection efficiency was counted by the average of three random fields of vision;cell viability was measured by MTT assay. Results 540 V/60μs/Pulsel/RT were found to be the optimal electric transfection conditions in HaCaT cell when the concentration of HaCaT cell and pGPU6/GFP/nev was 1.0 X 106 cells/ml and 2.0μg/mL. HaCaT cell had a better cellular growth behavior,and electrotransfection rate and cell survival rate were about 33.23% and 61.22%,respectively. Conclusion By optimizing the electransfection conditions,the electransfection rate of HaCaT cell is efficiently improved,which will lay the foundation for further studying gene functions.
ACCESSION #
97028819

 

Related Articles

Share

Read the Article

Courtesy of THE LIBRARY OF VIRGINIA

Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics