title: Activin-SMAD signaling is required for maintenance of porcine iPS cell self-renewal through upregulation of NANOG and OCT4 expression.
abstract: Porcine induced pluripotent stem cells (piPSCs) retain the enormous potential for farm animal reproduction and translational medicine, and have been reported by many laboratories worldwide. Some piPSC lines were bFGF-dependence and showed mouse EpiSC-like morphology; other lines were LIF-dependence and showed mouse ESC-like morphology. Metastable state of piPSC line that required both LIF and bFGF was also reported. Because bona fide pig embryonic stem cells were not available, uncovering piPSC state-specific regulatory circuitries was the most important task. In this study, we explored the function of Activin-SMAD signaling pathway and its downstream activated target genes in piPSCs. Transcriptome analysis showed that genes involved in Activin-SMAD signaling pathway were evidently activated during porcine somatic cell reprogramming, regardless piPSCs were LIF- or bFGF-dependent. Addition of Activin A and overexpression of SMAD2/3 significantly promoted expressions of porcine NANOG and OCT4, whereas inhibition of Activin-SMAD signaling by SB431542 and SMAD7 reduced NANOG and OCT4 expressions, and induced piPSCs differentiation exiting from pluripotent state. Our data demonstrate that activation of Activin-SMAD signaling pathway by addition of Activin A in culture medium is necessary for maintenance of self-renewal in porcine pluripotent stem cells.
authors.affiliation: Department of Animal Biotechnology, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
authors.affiliation: Department of Animal Biotechnology, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Wang
authors.affiliation: Department of Animal Biotechnology, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: Yaxian
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Animal Biotechnology, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: Tong
authors.initials: T
authors.lastname: Yu
authors.affiliation: Department of Animal Biotechnology, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: Huayan
authors.initials: H
authors.lastname: Wang
mesh_terms: D028341:Activins / Q000494:pharmacology*; D000818:Animals; D000066673:Cell Self Renewal / Q000187:drug effects*; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000187:drug effects; D000067470:Cellular Reprogramming Techniques; D018920:Coculture Techniques; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects* / Q000378:metabolism; D008813:Mice, Inbred ICR; D000071317:Nanog Homeobox Protein / Q000235:genetics / Q000378:metabolism*; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D051785:Smad Proteins / Q000235:genetics / Q000378:metabolism*; D051899:Smad2 Protein / Q000378:metabolism; D051900:Smad3 Protein / Q000378:metabolism; D051906:Smad7 Protein / Q000378:metabolism; D013552:Swine; D014162:Transfection; D015854:Up-Regulation
chemical_list: D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; D051785:Smad Proteins; D051899:Smad2 Protein; D051900:Smad3 Protein; D051906:Smad7 Protein; C422405:activin A; D028341:Activins
keywords: Activin-SMAD signaling; NANOG; iPS cells; pig; pluripotency