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Published on: December 28, 2015
[The expression level of the transfected hEGF gene to cultured human epidermal cells]
1Center of Burn, Changhai Hospital, Second Military Medical University, Shanghai 200433, PR China.
Objective:
To study the stable expression levels of cultured human epidermal cells transfected with a hEGF-plasmid.
Methods:
The 60%-70% confluent cultured human epidermal cells were transfected with a pBK-Signal-EGF plasmid-construct mediated by lipofectAMINE. After screening of G418 successfully transfected, clone forming cells were picked and subcultured for several weeks. To detect hEGF expression levels and to determine long term transgenic stability a specific ELISA was used.
Results:
(1) hEGF plasmid constructs were successfully introduced into cultured human epidermal cells. (2) Transgenic expression still persisted after four subcultures. (3) hEGF expression levels could be detected in the medium over a six-week period.
Conclusions:
The stable expression of hEGF was obtained after gene transfected with the pBK-Signal-EGF plasmid mediated by lipofectAMINE.
