Expression of Six Proteins Causes Reprogramming of Porcine Fibroblasts Into Induced Pluripotent Stem Cells With Both Active X Chromosomes |
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| Authors: | Tomokazu Fukuda Tetsuya Tani Seiki Haraguchi Kenichiro Donai Nobuyoshi Nakajima Hirohide Uenishi Takahiro Eitsuka Makoto Miyagawa Sanghoun Song Manabu Onuma Yumi Hoshino Eimei Sato Arata Honda |
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| Affiliation: | 1. United Graduate School of Agricultural Sciences, Iwate University, Morioka 020‐8551, Iwate, Japan;2. Laboratory of Animal Reproduction, Department of Advanced Bioscience, Faculty of Agriculture, Kindai University, Nara 631‐8505, Japan;3. Division of Animal Sciences, Animal Biotechnology Unit, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Tsukuba, Ibaraki 305‐0901, Japan;4. Graduate School of Agricultural Science, Tohoku University, Sendai, Japan;5. Center for Environmental Biology and Ecosystem Studies, National Institute of Environmental Studies, Tsukuba, Japan;6. Animal Bioregulation Unit, Division of Animal Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Tsukuba, Ibaraki 305‐8634, Japan;7. Faculty of Applied Life Sciences, Niigata University of Pharmacy and Applied Life Sciences, Niigata, Japan;8. Central Experimental Animal Center, Teikyo University School of Medicine, Japan;9. Faculty of Life and Environmental Science, Shimane University, Matsue, Shimane, Japan;10. Laboratory of Reproductive Endocrinology, Graduate School of Biosphere Science, Hiroshima University, Hiroshima 739‐8528, Japan;11. National Livestock Breeding Center, Fukushima 961‐8511, Japan;12. Organization for Promotion of Tenure Track, University of Miyazaki, Kiyotake, Miyazaki, Japan |
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| Abstract: | In this study, we created porcine‐induced pluripotent stem (iPS) cells with the expression of six reprogramming factors (Oct3/4, Klf4, Sox2, c‐Myc, Lin28, and Nanog). The resulting cells showed growth dependent on LIF (leukemia inhibitory factor) and expression of multiple stem cell markers. Furthermore, the iPS cells caused teratoma formation with three layers of differentiation and had both active X chromosomes (XaXa). Our iPS cells satisfied the both of important characteristics of stem cells: teratoma formation and activation of both X chromosomes. Injection of these iPS cells into morula stage embryos showed that these cells participate in the early stage of porcine embryogenesis. Furthermore, the RNA‐Seq analysis detected that expression levels of endogenous pluripotent related genes, NANOG, SOX2, ZFP42, OCT3/4, ESRRB, and ERAS were much higher in iPS with six factors than that with four reprogramming factors. We can conclude that the expression of six reprogramming factors enables the creation of porcine iPS cells, which is partially close to naive iPS state. J. Cell. Biochem. 118: 537–553, 2017. © 2016 Wiley Periodicals, Inc. |
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| Keywords: | PLURIPOTENT STEM CELL PORCINE X CHROMOSOME EPIGENETIC STATUS TRANSCRIPTIONAL FACTORS |
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