Porcine nuclear transfer using somatic donor cells altered to express male germ cell function
Sangho Roh A C , Hye-Yeon Choi B , Sang Kyu Park A , Cheolhee Won A , Bong-Woo Kim B , Jung-Hyun Kim B , Hoin Kang A , Eung-Ryoung Lee B and Ssang-Goo Cho B C
+ Author Affiliations
- Author Affiliations
A Embryo Biotechnology Laboratory, Dental Research Institute and CLS21, Seoul National University School of Dentistry, Seoul 110-749, Korea.
B Department of Animal Biotechnology (BK21) and RCTCP (CNU), Konkuk University, Seoul 143-701, Korea.
C Corresponding authors. Email: sangho@snu.ac.kr; ssangoo@konkuk.ac.kr
Reproduction, Fertility and Development 21(7) 882-891 https://doi.org/10.1071/RD09063
Submitted: 18 March 2009 Accepted: 17 May 2009 Published: 27 July 2009
Abstract
Recent studies reported that the direct transformation of one differentiated somatic cell type into another is possible. In the present study, we were able to modulate the cell fate of somatic cells to take on male germ cell function by introducing cell extracts derived from porcine testis tissue. Fibroblasts were treated with streptolysin O, which reversibly permeabilises the plasma membrane, and incubated with testis extracts. Our results showed that the testis extracts (TE) could activate expression of male germ cell-specific genes, implying that TE can provide regulatory components required for altering the cell fate of fibroblasts. Male germ cell function was sustained for more than 10 days after the introduction of TE. In addition, a single TE-treated cell was injected directly into the cytoplasm of in vitro-matured porcine oocytes. The rate of blastocyst formation was significantly higher in the TE-treated nuclear donor cell group than in the control cell group. The expression level of Nanog, Sox9 and Eomes was drastically increased when altered cells were used as donor nuclei. Our results suggest that TE can be used to alter the cell fate of fibroblasts to express male germ cell function and improve the developmental efficiency of the nuclear transfer porcine embryos.
Additional keywords: reprogramming, testis extracts.
Acknowledgements
We thank Dr H. K. Kim (Korea University) for kindly providing the stable transformed porcine cell line, V6 cells. This work was supported by the grant of ARPC (Grant no. 204117–03–3-CG000 for S. Roh and S.-G. Cho) in Korea and was also supported by the grants of KOSEF (ERC-R11–2002–100–05002–0 for S.-G. Cho and M10641000001–06N4100–00110 for S. Roh) in Korea.
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