THE CONSTRUCTION OF TRANSIENT GENE EXPRESSION TRANSFECTION SYSTEM IN MCF-7 CELLS
- VernacularTitle:MCF-7细胞瞬时基因表达转染模式的建立
- Author:
Haijiang ZHANG
;
Shiwei HAN
;
Jianli SANG
;
Dacheng HE
- Publication Type:Journal Article
- Keywords:
Cell transfection;
Transient gene expression;
Polyethylenimine(PEI)
- From:
Acta Anatomica Sinica
1954;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish an efficient, simple, low cytotoxicity and cheap transfaction system. Methods We have used the cationic polymer polyethylenimine(PEI) to study transient transfection in MCF-7 cells by testing different conditions, including cell concentrations, DNA concentrations, the ratio of PEI nitrogen to DNA phosphate(PEI-N∶DNA-P) and the time of cells grown in serum-free culture together with PEI-DNA complex. Results The optimized cell concentrations were 2?10 5 cells seeded per well in 24-well dishes 18-24?h before transfection. The DNA concentrations and ratio of PEI-N∶DNA-P are very important for optimal transfection and they affect each other. For 1??g DNA per well, the optimal PEI-N∶DNA-P is about 33∶1, however, as for 4??g DNA, it is 9∶1. The best time of cells grown in serum-free culture together with PEI-DNA complex is about 5-7?h.Conclusion With optimized conditions, we can establish an efficient, simple, low cytotoxicity and cheap transfection system by using PEI.
