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Journal of Animal and Veterinary Advances

Year: 2013  |  Volume: 12  |  Issue: 11  |  Page No.: 1032 - 1037

Immortalization of Swine Tracheal Epithelial Cells with Human Telomerase Reverse Transcriptase

Bao-Yu Chen, Kang-Kang Guo, Jin-Jin Wang, Lei He and Yan-Ming Zhang

Abstract

The tracheal epithelium is an important barrier that protects against harmful inhaled substances. To facilitate understanding of the mechanisms underlying respiratory diseases including tracheal cancer and tracheitis, researchers established an immortalized tracheal epithelial cell line. Primary cultures of Swine Tracheal Epithelial Cells (STECs) were immortalized by transfection of human Telomerase Reverse Transcriptase (hTERT; pCI-neo-hTERT) using lipofection. Positive cells were selected with G418 and expanded for continuous culture for up to 60 passages. The expression of hTERT mRNA in transfected cells was detected by RT-PCR. Transfected cells were assessed for morphology, karyotype, growth in soft agar and tumorigenicity in nude mice. Immortalized cells showed similar properties to those of normal cells such as contact inhibition, serum requirement and anchorage dependence. A soft agar assay and karyotype analysis showed no neoplastic transformation. These results suggest that immortalized STECs induced by the hTERT gene retain their original characteristics. The immortal STEC line may be useful as an in vitro model of tracheal epithelium for physiological, pathological and pharmacological investigations.

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