Immortalization of embryonic fibroblasts in heat shock transcription factor 1 knockout mouse.
- Author:
Mei-dong LIU
1
;
Hua-li ZHANG
;
Huan-yu GONG
;
Guang-wen CHEN
;
Kang-kai WANG
;
E SHUN-MEI
;
Xian-zhong XIAO
Author Information
1. Department of Pathophysiology, Xiangya School of Medicine, Central South University, Changsha, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antigens, Polyomavirus Transforming;
pharmacology;
Cell Line;
DNA-Binding Proteins;
genetics;
Embryo, Mammalian;
Female;
Fibroblasts;
cytology;
Heat Shock Transcription Factors;
Male;
Mice;
Mice, Knockout;
Transcription Factors;
genetics
- From:
Journal of Central South University(Medical Sciences)
2006;31(2):174-177
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To establish immortalized embryonic fibroblast lines in heat shock transcription factor 1 (HSF1) HSF1-/- and HSF1+/+ mice and to provide experimental models to study the function of HSF1.
METHODS:A mammalian expression vector (pSV3neo) containing the SV40 large T antigen was used to transfect the HSF1-/- and HSF1+/+ mouse embryonic fibroblast using Lipofectamine 2000. Colonies were screened by G418 and expanded to immortalized cell lines. PCR was used to detect the integration of the large T antigen with genome in the mouse embryonic fibroblast. Expression of SV40 large T antigen gene in expanded cells was identified by RT-PCR. HSP70 expression was examined by Western blot in the embryonic fibroblast lines.
RESULTS:The stable growth and serial propagation were observed in the HSF1-/- and HSF1+/+ cell lines for six months. The mRNA of SV40 T antigen gene expressed in the two cell lines. HSP70 expression could not be induced in the heat-treated HSF1-/- mouse embryo fibroblasts.
CONCLUSION:The immortalized cells of HSF1+/+ and HSF1-/- mouse embryo fibroblasts are successfully established.
