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Establishment and characterization of embryonic stem-like cells from porcine somatic cell nuclear transfer blastocysts

Published online by Cambridge University Press:  22 March 2010

S. Kim ,
J.H. Kim ,
E. Lee ,
Y.W. Jeong ,
M.S. Hossein ,
S.M. Park ,
S.W. Park ,
J.Y. Lee ,
Y.I. Jeong  and
H.S. Kim
...Show all authors
S. Kim
Affiliation:
SooAm Biotech Research Foundation, 1024–39 Saam-ri, Wonsam-myeon, Cheoin-gu, Yongin-si, Gyeonggi-do, 449–872, South Korea.
J.H. Kim
Affiliation:
SooAm Biotech Research Foundation, 1024–39 Saam-ri, Wonsam-myeon, Cheoin-gu, Yongin-si, Gyeonggi-do, 449–872, South Korea.
E. Lee
Affiliation:
SooAm Biotech Research Foundation, 1024–39 Saam-ri, Wonsam-myeon, Cheoin-gu, Yongin-si, Gyeonggi-do, 449–872, South Korea.
Y.W. Jeong
Affiliation:
SooAm Biotech Research Foundation, 1024–39 Saam-ri, Wonsam-myeon, Cheoin-gu, Yongin-si, Gyeonggi-do, 449–872, South Korea.
M.S. Hossein
Affiliation:
SooAm Biotech Research Foundation, 1024–39 Saam-ri, Wonsam-myeon, Cheoin-gu, Yongin-si, Gyeonggi-do, 449–872, South Korea.
S.M. Park
Affiliation:
SooAm Biotech Research Foundation, 1024–39 Saam-ri, Wonsam-myeon, Cheoin-gu, Yongin-si, Gyeonggi-do, 449–872, South Korea.
S.W. Park
Affiliation:
SooAm Biotech Research Foundation, 1024–39 Saam-ri, Wonsam-myeon, Cheoin-gu, Yongin-si, Gyeonggi-do, 449–872, South Korea.
J.Y. Lee
Affiliation:
SooAm Biotech Research Foundation, 1024–39 Saam-ri, Wonsam-myeon, Cheoin-gu, Yongin-si, Gyeonggi-do, 449–872, South Korea.
Y.I. Jeong
Affiliation:
SooAm Biotech Research Foundation, 1024–39 Saam-ri, Wonsam-myeon, Cheoin-gu, Yongin-si, Gyeonggi-do, 449–872, South Korea.
H.S. Kim
Affiliation:
SooAm Biotech Research Foundation, 1024–39 Saam-ri, Wonsam-myeon, Cheoin-gu, Yongin-si, Gyeonggi-do, 449–872, South Korea.
Y.W. Kim
Affiliation:
SooAm Biotech Research Foundation, 1024–39 Saam-ri, Wonsam-myeon, Cheoin-gu, Yongin-si, Gyeonggi-do, 449–872, South Korea.
S.H. Hyun
Affiliation:
SooAm Biotech Research Foundation, 1024–39 Saam-ri, Wonsam-myeon, Cheoin-gu, Yongin-si, Gyeonggi-do, 449–872, South Korea. Laboratory of Veterinary Biotechnology, College of Veterinary Medicine, Chungbuk National University, 410 Sungbong-ro, Heungduk-gu, Chungbuk, 361–763, South Korea.
W.S. Hwang*
Affiliation:
SooAm Biotech Research Foundation, 1024–39 Saam-ri, Wonsam-myeon, Cheoin-gu, Yongin-si, Gyeonggi-do, 449–872, South Korea.
*
All correspondence to: W.S. Hwang, SooAm Biotech Research Foundation, 1024–39 Saam-ri, Wonsam-myeon, Cheoin-gu, Yongin-si, Gyeonggi-do, 449–872, South Korea. Tel: +82 31 336 0500. Fax: +82 31 336 8439. e-mail: hwangws@sooam.org
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Summary

This study was aimed to establish embryonic stem (ES)-like cells from blastocysts derived from somatic cell nuclear transfer (SCNT) in pig. Somatic cells isolated from both day-30 fetus and neonatal cloned piglet were used for donor cells. A total of 60 blastocysts (46 and 14 derived from fetal and neonatal fibroblast donor cells, respectively) were seeded onto a mitotically inactive mouse embryonic fibroblast (MEF) monolayer and two ES-like cell lines, one from each donor cell type, were established. They remained undifferentiated over more than 52 (fetal fibroblast-derived) and 48 (neonatal fibroblast-derived) passages, while retaining alkaline phosphatase activity and reactivity with ES specific markers Oct-4, stage-specific embryonic antigen-1 (SSEA-1), SSEA-4, TRA-1–60 and TRA-1–81. These ES-like cells maintained normal diploid karyotype throughout subculture and successfully differentiated into embryoid bodies that expressed three germ layer-specific genes (ectoderm: β-III tubulin; endoderm: amylase; and mesoderm: enolase) after culture in leukemia inhibitory factor-free medium. Microsatellite analysis confirmed that they were genetically identical to its donor cells. Combined with gene targeting, our results may contribute to developing an efficient method for producing transgenic pigs for various purposes.

Keywords

Embryonic stem (ES)-like cellsSomatic cell nuclear transferTransgenic pigs
Type
Research Article
Information
Zygote , Volume 18 , Issue 2 , May 2010 , pp. 93 - 101
Copyright
Copyright © Cambridge University Press 2010

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