Efficient establishment of pig embryonic fibroblast cell lines with conditional expression of the simian vacuolating virus 40 large T fragment

Tomokazu Fukuda; Masafumi Katayama; Takayuki Yoshizawa; Takahiro Eitsuka; Hiroaki Mizukami; Kiyotaka Nakagawa; Hisao Ito; Homika Komagata; Sanghoun Song; Sanggun Roh; Yumi Hoshino; Eimei Sato; Hirofumi Hanada; Katsuhiko Nishimori; Teruo Miyazawa; Takafumi Uchida

doi:10.1271/bbb.120155

Efficient establishment of pig embryonic fibroblast cell lines with conditional expression of the simian vacuolating virus 40 large T fragment

Tomokazu Fukuda, Masafumi Katayama, Takayuki Yoshizawa, Takahiro Eitsuka, Hiroaki Mizukami, Kiyotaka Nakagawa, Hisao Ito, Homika Komagata, Sanghoun Song, Sanggun Roh, Yumi Hoshino, Eimei Sato, Hirofumi Hanada, Katsuhiko Nishimori, Teruo Miyazawa, Takafumi Uchida

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20 Citations (Scopus)

Abstract

The pig is an important animal for both agricultural and medical purposes. However, the number of pig-derived cell lines is relatively limited when compared with mouse-and human-derived lines. We established in this study a retroviral conditional expression system for the Simian vacuolating virus 40 large T fragment (SV40T) which allowed us to efficiently establish pig embryonic fibroblast cell lines. The established cell lines showed high levels of cell proliferation and resistance to cellular senescence. A chromosome analysis showed that 84% of the cells had the normal karyotype. Transient expression of the Cre recombinase allowed us to excise the SV40T fragment from the genome. The development of this research tool will enable us to quickly establish new cell lines derived from various animals.

Original language	English
Pages (from-to)	1372-1377
Number of pages	6
Journal	Bioscience, Biotechnology and Biochemistry
Volume	76
Issue number	7
DOIs	https://doi.org/10.1271/bbb.120155
Publication status	Published - 2012

Keywords

Conditional expression system
Immortalization
Retrovirus
Simian vacuolating virus 40 large T fragment (SV40T)

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10.1271/bbb.120155

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Fukuda, T., Katayama, M., Yoshizawa, T., Eitsuka, T., Mizukami, H., Nakagawa, K., Ito, H., Komagata, H., Song, S., Roh, S., Hoshino, Y., Sato, E., Hanada, H., Nishimori, K., Miyazawa, T., & Uchida, T. (2012). Efficient establishment of pig embryonic fibroblast cell lines with conditional expression of the simian vacuolating virus 40 large T fragment. Bioscience, Biotechnology and Biochemistry, 76(7), 1372-1377. https://doi.org/10.1271/bbb.120155

@article{848dbb4a26dd48f0a3a6e16cd1588dae,

title = "Efficient establishment of pig embryonic fibroblast cell lines with conditional expression of the simian vacuolating virus 40 large T fragment",

abstract = "The pig is an important animal for both agricultural and medical purposes. However, the number of pig-derived cell lines is relatively limited when compared with mouse-and human-derived lines. We established in this study a retroviral conditional expression system for the Simian vacuolating virus 40 large T fragment (SV40T) which allowed us to efficiently establish pig embryonic fibroblast cell lines. The established cell lines showed high levels of cell proliferation and resistance to cellular senescence. A chromosome analysis showed that 84% of the cells had the normal karyotype. Transient expression of the Cre recombinase allowed us to excise the SV40T fragment from the genome. The development of this research tool will enable us to quickly establish new cell lines derived from various animals.",

keywords = "Conditional expression system, Immortalization, Retrovirus, Simian vacuolating virus 40 large T fragment (SV40T)",

author = "Tomokazu Fukuda and Masafumi Katayama and Takayuki Yoshizawa and Takahiro Eitsuka and Hiroaki Mizukami and Kiyotaka Nakagawa and Hisao Ito and Homika Komagata and Sanghoun Song and Sanggun Roh and Yumi Hoshino and Eimei Sato and Hirofumi Hanada and Katsuhiko Nishimori and Teruo Miyazawa and Takafumi Uchida",

note = "Funding Information: This study was supported by KAKENHI, Grants-in-Aid for Scientific Research from Japan Society for the Promotion of Science, a research grant from the Suzuken Memorial Foundation, and a research grant from the Iijima Memorial Foundation for the Promotion of Food, Science, and Technology. We thank Dr. Masahito Ikawa (Osaka University) for kindly providing the D64V mutant packaging plasmid.",

year = "2012",

doi = "10.1271/bbb.120155",

language = "English",

volume = "76",

pages = "1372--1377",

journal = "Bioscience, Biotechnology and Biochemistry",

issn = "0916-8451",

publisher = "Oxford University Press",

number = "7",

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Fukuda, T, Katayama, M, Yoshizawa, T, Eitsuka, T, Mizukami, H, Nakagawa, K, Ito, H, Komagata, H, Song, S, Roh, S, Hoshino, Y, Sato, E, Hanada, H, Nishimori, K, Miyazawa, T & Uchida, T 2012, 'Efficient establishment of pig embryonic fibroblast cell lines with conditional expression of the simian vacuolating virus 40 large T fragment', Bioscience, Biotechnology and Biochemistry, vol. 76, no. 7, pp. 1372-1377. https://doi.org/10.1271/bbb.120155

TY - JOUR

T1 - Efficient establishment of pig embryonic fibroblast cell lines with conditional expression of the simian vacuolating virus 40 large T fragment

AU - Fukuda, Tomokazu

AU - Katayama, Masafumi

AU - Yoshizawa, Takayuki

AU - Eitsuka, Takahiro

AU - Mizukami, Hiroaki

AU - Nakagawa, Kiyotaka

AU - Ito, Hisao

AU - Komagata, Homika

AU - Song, Sanghoun

AU - Roh, Sanggun

AU - Hoshino, Yumi

AU - Sato, Eimei

AU - Hanada, Hirofumi

AU - Nishimori, Katsuhiko

AU - Miyazawa, Teruo

AU - Uchida, Takafumi

N1 - Funding Information: This study was supported by KAKENHI, Grants-in-Aid for Scientific Research from Japan Society for the Promotion of Science, a research grant from the Suzuken Memorial Foundation, and a research grant from the Iijima Memorial Foundation for the Promotion of Food, Science, and Technology. We thank Dr. Masahito Ikawa (Osaka University) for kindly providing the D64V mutant packaging plasmid.

PY - 2012

Y1 - 2012

N2 - The pig is an important animal for both agricultural and medical purposes. However, the number of pig-derived cell lines is relatively limited when compared with mouse-and human-derived lines. We established in this study a retroviral conditional expression system for the Simian vacuolating virus 40 large T fragment (SV40T) which allowed us to efficiently establish pig embryonic fibroblast cell lines. The established cell lines showed high levels of cell proliferation and resistance to cellular senescence. A chromosome analysis showed that 84% of the cells had the normal karyotype. Transient expression of the Cre recombinase allowed us to excise the SV40T fragment from the genome. The development of this research tool will enable us to quickly establish new cell lines derived from various animals.

AB - The pig is an important animal for both agricultural and medical purposes. However, the number of pig-derived cell lines is relatively limited when compared with mouse-and human-derived lines. We established in this study a retroviral conditional expression system for the Simian vacuolating virus 40 large T fragment (SV40T) which allowed us to efficiently establish pig embryonic fibroblast cell lines. The established cell lines showed high levels of cell proliferation and resistance to cellular senescence. A chromosome analysis showed that 84% of the cells had the normal karyotype. Transient expression of the Cre recombinase allowed us to excise the SV40T fragment from the genome. The development of this research tool will enable us to quickly establish new cell lines derived from various animals.

KW - Conditional expression system

KW - Immortalization

KW - Retrovirus

KW - Simian vacuolating virus 40 large T fragment (SV40T)

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AN - SCOPUS:84864274949

SN - 0916-8451

VL - 76

SP - 1372

EP - 1377

JO - Bioscience, Biotechnology and Biochemistry

JF - Bioscience, Biotechnology and Biochemistry

IS - 7

ER -

Efficient establishment of pig embryonic fibroblast cell lines with conditional expression of the simian vacuolating virus 40 large T fragment

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Keywords

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