title: Systematic in vitro and in vivo characterization of Leukemia-inhibiting factor- and Fibroblast growth factor-derived porcine induced pluripotent stem cells.
abstract: Derivation and stable maintenance of porcine induced pluripotent stem cells (piPSCs) is challenging. We herein systematically analyzed two piPSC lines, derived by lentiviral transduction and cultured under either leukemia inhibitory factor (LIF) or fibroblast growth factor (FGF) conditions, to shed more light on the underlying biological mechanisms of porcine pluripotency. LIF-derived piPSCs were more successful than their FGF-derived counterparts in the generation of in vitro chimeras and in teratoma formation. When LIF piPSCs chimeras were transferred into surrogate sows and allowed to develop, only their prescence within the embryonic membranes could be detected. Whole-transcriptome analysis of the piPSCs and porcine neonatal fibroblasts showed that they clustered together, but apart from the two pluripotent cell populations of early porcine embryos, indicating incomplete reprogramming. Indeed, bioinformatic analysis of the pluripotency-related gene network of the LIF- versus FGF-derived piPSCs revealed that ZFP42 (REX1) expression was absent in both piPSC-like cells, whereas it was expressed in the porcine inner cell mass at Day 7/8. A second striking difference was the expression of ATOH1 in piPSC-like cells, which was absent in the inner cell mass. Moreover, our gene expression analyses plus correlation analyses of known pluripotency genes identified unique relationships between pluripotency genes in the inner cell mass, which are to some extent, in the piPSC-like cells. This deficiency in downstream gene activation and divergent gene expression may be underlie the inability to derive germ line-transmitting piPSCs, and provides unique insight into which genes are necessary to achieve fully reprogrammed piPSCs. 84: 229-245, 2017. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Veterinary Reproduction and Obstetrics, Faculty of Health and Medical Sciences, Department of Large Animal Sciences, University of Copenhagen, Frederiksberg C, Denmark.
authors.forename: Jan O
authors.initials: JO
authors.lastname: Secher
authors.affiliation: Faculty of Veterinary Medicine Ankara University, Department of Histology and Embryology, Diskapi, Ankara, Turkey.
authors.forename: Ahmet
authors.initials: A
authors.lastname: Ceylan
authors.affiliation: Animal Breeding, Quantitative Genetics and Systems Biology Group, Faculty of Health and Medical Sciences, Department of Large Animal Sciences, University of Copenhagen, Frederiksberg C, Denmark.
authors.forename: Gianluca
authors.initials: G
authors.lastname: Mazzoni
authors.affiliation: Faculty of Health and Medical Sciences, Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, Frederiksberg C, Denmark.|BioTalentum Ltd., Gödöllő, Hungary.|Faculty of Veterinary Medicine, Departments of Equine Sciences and Farm Animal Health, Utrecht University, Utrecht, Netherlands.
authors.forename: Kaveh
authors.initials: K
authors.lastname: Mashayekhi
authors.affiliation: Faculty of Health and Medical Sciences, Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, Frederiksberg C, Denmark.|BioTalentum Ltd., Gödöllő, Hungary.|Faculty of Veterinary Medicine, Departments of Equine Sciences and Farm Animal Health, Utrecht University, Utrecht, Netherlands.
authors.forename: Tong
authors.initials: T
authors.lastname: Li
authors.affiliation: BioTalentum Ltd., Gödöllő, Hungary.|Faculty of Veterinary Medicine, Department of Farm Animal Health, Utrecht University, Utrecht, Netherlands.
authors.forename: Suchitra
authors.initials: S
authors.lastname: Muenthaisong
authors.affiliation: Danish Dementia Research Centre, Rigshospitalet, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Troels T
authors.initials: TT
authors.lastname: Nielsen
authors.affiliation: Faculty of Health and Medical Sciences, Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, Frederiksberg C, Denmark.
authors.forename: Dong
authors.initials: D
authors.lastname: Li
authors.affiliation: Department of Biomedicine, Aarhus University, Aarhus C, Denmark.
authors.forename: Shengting
authors.initials: S
authors.lastname: Li
authors.affiliation: Institute for Farm Animal Genetics (FLI), Neustadt, Germany.
authors.forename: Stoyan
authors.initials: S
authors.lastname: Petkov
authors.affiliation: Faculty of Health and Medical Sciences, Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, Frederiksberg C, Denmark.
authors.forename: Susanna
authors.initials: S
authors.lastname: Cirera
authors.affiliation: Department of Biomedicine, Aarhus University, Aarhus C, Denmark.
authors.forename: Yonglun
authors.initials: Y
authors.lastname: Luo
authors.affiliation: Department of Statistics, University of Missouri, Columbia, Missouri.
authors.forename: Lori
authors.initials: L
authors.lastname: Thombs
authors.affiliation: Animal Breeding, Quantitative Genetics and Systems Biology Group, Faculty of Health and Medical Sciences, Department of Large Animal Sciences, University of Copenhagen, Frederiksberg C, Denmark.
authors.forename: Haja N
authors.initials: HN
authors.lastname: Kadarmideen
authors.affiliation: BioTalentum Ltd., Gödöllő, Hungary.|Faculty of Veterinary Medicine, Departments of Equine Sciences and Farm Animal Health, Utrecht University, Utrecht, Netherlands.|Molecular Animal Biotechnology Laboratory, Szent István University, Gödöllő, Hungary.
authors.forename: Andras
authors.initials: A
authors.lastname: Dinnyes
authors.affiliation: Department of Biomedicine, Aarhus University, Aarhus C, Denmark.
authors.forename: Lars
authors.initials: L
authors.lastname: Bolund
authors.affiliation: Faculty of Veterinary Medicine, Department of Farm Animal Health, Utrecht University, Utrecht, Netherlands.
authors.forename: Bernard A J
authors.initials: BA
authors.lastname: Roelen
authors.affiliation: Veterinary Reproduction and Obstetrics, Faculty of Health and Medical Sciences, Department of Large Animal Sciences, University of Copenhagen, Frederiksberg C, Denmark.
authors.forename: Mette
authors.initials: M
authors.lastname: Schmidt
authors.affiliation: Department of Animal Science, Aarhus University, Tjele, Denmark.
authors.forename: Henrik
authors.initials: H
authors.lastname: Callesen
authors.affiliation: Faculty of Health and Medical Sciences, Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, Frederiksberg C, Denmark.
authors.forename: Poul
authors.initials: P
authors.lastname: Hyttel
authors.affiliation: Faculty of Health and Medical Sciences, Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, Frederiksberg C, Denmark.
authors.forename: Kristine K
authors.initials: KK
authors.lastname: Freude
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism*; D005346:Fibroblast Growth Factors / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects*; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D051741:Kruppel-Like Transcription Factors / Q000378:metabolism*; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology*; D013552:Swine
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D005346:Fibroblast Growth Factors; D051741:Kruppel-Like Transcription Factors; D053605:Leukemia Inhibitory Factor