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Establishment and characterization of a telomerase‐immortalized porcine bronchial epithelial cell line
Author(s) -
Xie Xing,
Gan Yuan,
Pang Maoda,
Shao Guoqing,
Zhang Lei,
Liu Beibei,
Xu Qi,
Wang Haiyan,
Feng Yanyan,
Yu Yanfei,
Chen Rong,
Wu Meng,
Zhang Zhenzhen,
Hua Lizhong,
Xiong Qiyan,
Liu Maojun,
Feng Zhixin
Publication year - 2018
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.26942
Subject(s) - telomerase reverse transcriptase , telomerase , immortalised cell line , biology , cell culture , microbiology and biotechnology , biochemistry , genetics , gene
Primary porcine bronchial epithelial cells (PBECs) are an ideal model to study the molecular and pathogenic mechanisms of various porcine respiratory pathogens. However, the short lifespan of primary PBECs greatly limit their application. Here, we isolated and cultured primary PBECs and established immortalized PBECs by transfecting primary PBECs with the pEGFP‐hTERT recombinant plasmid containing human telomerase reverse transcriptase (hTERT). Immortalized PBECs (hTERT‐PBECs) retained the morphological and functional features of primary PBECs as indicated by cytokeratin 18 expression, telomerase activity assay, proliferation assays, karyotype analysis, and quantitative reverse‐transcriptase polymerase chain reaction. Compared to primary PBECs, hTERT‐PBECs had higher telomerase activity, extended replicative lifespan, and displayed enhanced proliferative activity. Moreover, this cell line is not transformed in vitro and does not exhibit a malignant phenotype in vivo, suggesting that it can be safely used in further studies. Besides, hTERT‐PBECs were susceptible to swine influenza virus of H3N2 subtype and porcine circovirus type 2. In conclusion, the immortalized hTERT‐PBECs represent a valuable in vitro model, which can be widely used in the study of porcine respiratory pathogenic infections.