Expression of Six Proteins Causes Reprogramming of Porcine Fibroblasts Into Induced Pluripotent Stem Cells With Both Active X Chromosomes.
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.
United Graduate School of Agricultural Sciences, Iwate University, 4-3-5, Ueda, Morioka 020-8551, Iwate, Japan.
Tomokazu
T
Fukuda
Laboratory of Animal Reproduction, Department of Advanced Bioscience, Faculty of Agriculture, Kindai University, 3327-204 Nakamachi, Nara 631-8505, Japan.
Tetsuya
T
Tani
Division of Animal Sciences, Animal Biotechnology Unit, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Tsukuba, Ibaraki 305-0901, Japan.
Seiki
S
Haraguchi
Graduate School of Agricultural Science, Tohoku University, Sendai, 981-8555, Japan.
Kenichiro
K
Donai
Center for Environmental Biology and Ecosystem Studies, National Institute of Environmental Studies, Tsukuba, Japan.
Nobuyoshi
N
Nakajima
Animal Bioregulation Unit, Division of Animal Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), 1-2 Owashi, Tsukuba, Ibaraki 305-8634, Japan.
Hirohide
H
Uenishi
Faculty of Applied Life Sciences, Niigata University of Pharmacy and Applied Life Sciences, Niigata, Japan.
Takahiro
T
Eitsuka
Central Experimental Animal Center, Teikyo University School of Medicine, Japan.
Makoto
M
Miyagawa
Faculty of Life and Environmental Science, Shimane University, Matsue, Shimane, Japan.
Sanghoun
S
Song
Center for Environmental Biology and Ecosystem Studies, National Institute of Environmental Studies, Tsukuba, Japan.
Manabu
M
Onuma
Laboratory of Reproductive Endocrinology, Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima, Kagamiyama 1-4-4, Hiroshima 739-8528, Japan.
Yumi
Y
Hoshino
National Livestock Breeding Center, Odakurahara, Odakura, Nishigo-mura, Nishishirakawa-gun, Fukushima 961-8511, Japan.
Eimei
E
Sato
Organization for Promotion of Tenure Track, University of Miyazaki, 5200 Kihara, Kiyotake, Miyazaki, 889-1692, Japan.
Arata
A
Honda
D000818:Animals; D000067470:Cellular Reprogramming Techniques; D005260:Female; D015870:Gene Expression; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism*; D013552:Swine; D014157:Transcription Factors / Q000096:biosynthesis*; D014960:X Chromosome / Q000378:metabolism*
D014157:Transcription Factors
EPIGENETIC STATUS; PLURIPOTENT STEM CELL; PORCINE; TRANSCRIPTIONAL FACTORS; X CHROMOSOME