Primed Pluripotent Cell Lines Derived from Various Embryonic Origins and Somatic Cells in Pig |
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Authors: | Jin-Kyu Park Hye-Sun Kim Kyung-Jun Uh Kwang-Hwan Choi Hyeong-Min Kim Taeheon Lee Byung-Chul Yang Hyun-Jong Kim Hak-Hyun Ka Heebal Kim Chang-Kyu Lee |
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Affiliation: | 1. Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute for Agriculture and Life Science, Seoul National University, Seoul, Republic of Korea.; 2. National Institute of Animal Science, RDA, Suwon, Republic of Korea.; 3. Department of Biological Resources and Technology, Yonsei University, Wonju, Republic of Korea.; University of Cincinnati, United States of America, |
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Abstract: | Since pluripotent embryonic stem cell (ESC) lines were first derived from the mouse, tremendous efforts have been made to establish ESC lines in several domestic species including the pig; however, authentic porcine ESCs have not yet been established. It has proven difficult to maintain an ESC-like state in pluripotent porcine cell lines due to the frequent occurrence of spontaneous differentiation into an epiblast stem cell (EpiSC)-like state during culture. We have been able to derive EpiSC-like porcine ESC (pESC) lines from blastocyst stage porcine embryos of various origins, including in vitro fertilized (IVF), in vivo derived, IVF aggregated, and parthenogenetic embryos. In addition, we have generated induced pluripotent stem cells (piPSCs) via plasmid transfection of reprogramming factors (Oct4, Sox2, Klf4, and c-Myc) into porcine fibroblast cells. In this study, we analyzed characteristics such as marker expression, pluripotency and the X chromosome inactivation status in female of our EpiSC-like pESC lines along with our piPSC line. Our results show that these cell lines demonstrate the expression of genes associated with the Activin/Nodal and FGF2 pathways along with the expression of pluripotent markers Oct4, Sox2, Nanog, SSEA4, TRA 1–60 and TRA 1–81. Furthermore all of these cell lines showed in vitro differentiation potential, the X chromosome inactivation in female and a normal karyotype. Here we suggest that the porcine species undergoes reprogramming into a primed state during the establishment of pluripotent stem cell lines. |
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