Establishment and characterization of a telomerase‐immortalized porcine bronchial epithelial cell line. Issue 12 (5th August 2018)

• Record Type: Journal Article
• Authors: 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
• Is Part Of: Journal of cellular physiology. Volume 233:Issue 12(2018:Dec.)
• Page Start: 9763
• Page End: 9776
• Publication Date: 2018-08-05
• Subjects: cell model -- hTERT‐PBECs -- porcine bronchial epithelial cells -- porcine respiratory disease; Physiology -- Periodicals; Cell physiology -- Periodicals; 571.6
• DOI: 10.1002/jcp.26942
• Languages: English
• ISSNs: 0021-9541

Abstract

Abstract : 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. Abstract : In summary, we provided detailed protocols for bronchus isolation from piglets and in vitro expansion of primary PBECs. Furthermore, we established the immortalized hTERT‐PBEC line, which retains the morphological and functional features of primary PBECs. The establishment of immortal hTERT‐PBECs is of great importance as an in vitro model for pathogenicity studies of porcine respiratory pathogens.