title: Derivation and characterization of sleeping beauty transposon-mediated porcine induced pluripotent stem cells.
abstract: The domestic pig is an important large animal model for preclinical testing of novel cell therapies. Recently, we produced pluripotency reporter pigs in which the Oct4 promoter drives expression of the enhanced green fluorescent protein (EGFP). Here, we reprogrammed Oct4-EGFP fibroblasts employing the nonviral Sleeping Beauty transposon system to deliver the reprogramming factors Oct4, Sox2, Klf4, and cMyc. Successful reprogramming to a pluripotent state was indicated by changes in cell morphology and reactivation of the Oct4-EGFP reporter. The transposon-reprogrammed induced pluripotent stem (iPS) cells showed long-term proliferation in vitro over >40 passages, expressed transcription factors typical of embryonic stem cells, including OCT4, NANOG, SOX2, REX1, ESRRB, DPPA5, and UTF1 and surface markers of pluripotency, including SSEA-1 and TRA-1-60. In vitro differentiation resulted in derivatives of the 3 germ layers. Upon injection of putative iPS cells under the skin of immunodeficient mice, we observed teratomas in 3 of 6 cases. These results form the basis for in-depth studies toward the derivation of porcine iPS cells, which hold great promise for preclinical testing of novel cell therapies in the pig model.
authors.affiliation: Institute of Farm Animal Genetics, Friedrich-Loeffler-Institut, Mariensee, Germany. wilfried.kues@fli.bund.de
authors.forename: Wilfried A
authors.initials: WA
authors.lastname: Kues
authors.forename: Doris
authors.initials: D
authors.lastname: Herrmann
authors.forename: Brigitte
authors.initials: B
authors.lastname: Barg-Kues
authors.forename: Srividiameena
authors.initials: S
authors.lastname: Haridoss
authors.forename: Monika
authors.initials: M
authors.lastname: Nowak-Imialek
authors.forename: Thomas
authors.initials: T
authors.lastname: Buchholz
authors.forename: Miriam
authors.initials: M
authors.lastname: Streeck
authors.forename: Antonia
authors.initials: A
authors.lastname: Grebe
authors.forename: Ivana
authors.initials: I
authors.lastname: Grabundzija
authors.forename: Sylvia
authors.initials: S
authors.lastname: Merkert
authors.forename: Ulrich
authors.initials: U
authors.lastname: Martin
authors.forename: Vanessa J
authors.initials: VJ
authors.lastname: Hall
authors.forename: Mikkel A
authors.initials: MA
authors.lastname: Rasmussen
authors.forename: Zoltan
authors.initials: Z
authors.lastname: Ivics
authors.forename: Poul
authors.initials: P
authors.lastname: Hyttel
authors.forename: Heiner
authors.initials: H
authors.lastname: Niemann
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism; D002471:Cell Transformation, Neoplastic; D002478:Cells, Cultured; D018920:Coculture Techniques; D004251:DNA Transposable Elements / Q000235:genetics*; D005347:Fibroblasts / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057026:Induced Pluripotent Stem Cells / Q000502:physiology* / Q000637:transplantation / Q000648:ultrastructure; D000090062:Kruppel-Like Factor 4; D051379:Mice; D008819:Mice, Nude; D008856:Microscopy, Fluorescence; D055495:Neurogenesis; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D034421:Sus scrofa; D013724:Teratoma / Q000473:pathology; D059467:Transcriptome; D019076:Transgenes
chemical_list: D000943:Antigens, Differentiation; D004251:DNA Transposable Elements; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D050814:Octamer Transcription Factor-3; D055748:SOXB1 Transcription Factors; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins