Establishment and characterization of a telomerase-immortalized canine bronchiolar epithelial cell line

Verfasser / Beitragende:
[Xing Xie, Maoda Pang, Shan Liang, Lei Yu, Yanbing Zhao, Ke Ma, Dildar Kalhoro, Chengping Lu, Yongjie Liu]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/21(2015-11-01), 9135-9146
Format:
Artikel (online)
ID: 605505837
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024 7 0 |a 10.1007/s00253-015-6794-8  |2 doi 
035 |a (NATIONALLICENCE)springer-10.1007/s00253-015-6794-8 
245 0 0 |a Establishment and characterization of a telomerase-immortalized canine bronchiolar epithelial cell line  |h [Elektronische Daten]  |c [Xing Xie, Maoda Pang, Shan Liang, Lei Yu, Yanbing Zhao, Ke Ma, Dildar Kalhoro, Chengping Lu, Yongjie Liu] 
520 3 |a Dogs are susceptible to infectious diseases that occur primarily in the respiratory tract. The airway epithelium acts as a first line of defense and is constantly exposed to microorganisms present in the environment. Respiratory epithelial cells have recently gained wide use as a cell model for studying the pathogenesis of human, murine or swine respiratory pathogen infections. However, studies of the pathogenic mechanisms of canine pathogens have been hindered by the lack of reliable respiratory cell lines. Here, we cultured primary canine bronchiolar epithelial cells (CBECs), whose characteristics were confirmed by their expression of the epithelial cell-specific marker cytokeratin 18, and have provided protocols for their isolation and ex vivo expansion. Further, we established immortalized CBECs containing the human telomerase reverse transcriptase (hTERT) gene via transfection of primary CBECs with the recombinant plasmid pEGFP-hTERT. Immortalized bronchiolar epithelial cells (hTERT-CBECs) retain the morphological and functional features of primary CBECs, as indicated by reverse transcriptase polymerase chain reaction, proliferation assays, karyotype analysis, telomerase activity assay, and Western blotting, which demonstrate that hTERT-CBECs have higher telomerase activity, an extended proliferative lifespan, and a diploid complement of chromosomes, even after Passage 50. Moreover, this cell line is not transformed, as evaluated using soft agar assays and tumorigenicity analysis in nude mice, and can therefore be safely used in future studies. The isolation and establishment of stable hTERT-CBECs is of great importance for use as an in vitro model for mechanistic studies of canine pathogenic infections. 
540 |a Springer-Verlag Berlin Heidelberg, 2015 
690 7 |a Canine bronchiolar epithelial cells (CBECs)  |2 nationallicence 
690 7 |a hTERT-CBECs  |2 nationallicence 
690 7 |a Pathogenesis  |2 nationallicence 
690 7 |a Cell model  |2 nationallicence 
700 1 |a Xie  |D Xing  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
700 1 |a Pang  |D Maoda  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
700 1 |a Liang  |D Shan  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
700 1 |a Yu  |D Lei  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
700 1 |a Zhao  |D Yanbing  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
700 1 |a Ma  |D Ke  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
700 1 |a Kalhoro  |D Dildar  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
700 1 |a Lu  |D Chengping  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
700 1 |a Liu  |D Yongjie  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/21(2015-11-01), 9135-9146  |x 0175-7598  |q 99:21<9135  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-015-6794-8  |q text/html  |z Onlinezugriff via DOI 
898 |a BK010053  |b XK010053  |c XK010000 
900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
908 |D 1  |a research-article  |2 jats 
949 |B NATIONALLICENCE  |F NATIONALLICENCE  |b NL-springer 
950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00253-015-6794-8  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Xie  |D Xing  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Pang  |D Maoda  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Liang  |D Shan  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Yu  |D Lei  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Zhao  |D Yanbing  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Ma  |D Ke  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Kalhoro  |D Dildar  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Lu  |D Chengping  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Liu  |D Yongjie  |u College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/21(2015-11-01), 9135-9146  |x 0175-7598  |q 99:21<9135  |1 2015  |2 99  |o 253