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Cell Reprogram. 2019 Feb;21(1):26-36. doi: 10.1089/cell.2018.0036.

Production of Transgenic Porcine Embryos Reconstructed with Induced Pluripotent Stem-Like Cells Derived from Porcine Endogenous Factors Using piggyBac System.

Author information

1
1 Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Research Institute of Veterinary Science, Seoul National University, Seoul, Republic of Korea.
2
2 ToolGen, Inc., Seoul, Republic of Korea.
3
3 BK21 Plus program, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea.
4
4 Emergence Center for Food-Medicine Personalized Therapy System, Advanced Institutes of Convergence Technology, Seoul National University, Gyeonggi-do, Republic of Korea.

Abstract

The potential of induced pluripotent stem (iPS) cells, which have self-renewal ability and can differentiate into three germ layers, led us to hypothesize that iPS cells in pigs can be useful and suitable source for producing transgenic pigs. In this study, we generated iPS-like cells using doxycycline-inducible piggyBac (PB) expression vectors encoding porcine 4 transcription factors. After transfection, transfected cells were cultured until the formation of outgrowing colonies taking least of 7-10 days. The iPS-like cells demonstrated pluripotent characteristics such as self-renewal, high proliferation, expression of pluripotent markers, and aggregation ability. The embryo development through somatic cell nuclear transfer (SCNT), cleavage rate, and blastocyst formation rate did not show any significant differences. However, the total cell number of blastocysts was significantly increased with the established cell line. In conclusion, the iPS-like cell line, generated from porcine transcriptional factors using the PB transposon system, demonstrated pluripotency with the capacity for unlimited self-renewal, and could be used as donor cells to produce cloned embryos by SCNT. These cells will be suitable for gene modification and would contribute to the stability or safety of pig models in biomedical research.

KEYWORDS:

transposon; donor cells; induced pluripotent cells; porcine transcription factors; somatic cell nuclear transfer

PMID:
30735078
DOI:
10.1089/cell.2018.0036

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