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Title of Journal: Cytotechnology

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Abbravation: Cytotechnology

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Publisher

Springer Netherlands

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DOI

10.1002/adma.201501934

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ISSN

1573-0778

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Immortalization of pig fibroblast cells by transpo

Authors: Shan He Yangyang Li Yang Chen Yue Zhu Xinyu Zhang Xiaoli Xia Huaichang Sun
Publish Date: 2015/09/04
Volume: 68, Issue: 4, Pages: 1435-1445
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Abstract

Pigs are the most economically important livestock but pig cell lines useful for physiological studies and/or vaccine development are limited Although several pig cell lines have been generated by oncogene transformation or human telomerase reverse transcriptase TERT immortalization these cell lines contain viral sequences and/or antibiotic resistance genes In this study we established a new method for generating pig cell lines using the Sleeping Beauty SB transposonmediated ectopic expression of porcine telomerase reverse transcriptase pTERT The performance of the new method was confirmed by generating a pig fibroblast cell PFC line After transfection of primary PFCs with the SB transposon system one cell clone containing the pTERT expression cassette was selected by dilution cloning and passed for different generations After passage for more than 40 generations the cell line retained stable expression of ectopic pTERT and continuous growth potential Further characterization showed that the cell line kept the fibroblast morphology growth curve population doubling time cloning efficiency marker gene expression pattern cell cycle distribution and anchoragedependent growth property of the primary cells These data suggest that the new method established is useful for generating pig cell lines without viral sequence and antibiotic resistant gene

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