title: Porcine induced pluripotent stem cells produce chimeric offspring.
abstract: Ethical and moral issues rule out the use of human induced pluripotent stem cells (iPSCs) in chimera studies that would determine the full extent of their reprogrammed state, instead relying on less rigorous assays such as teratoma formation and differentiated cell types. To date, only mouse iPSC lines are known to be truly pluripotent. However, initial mouse iPSC lines failed to form chimeric offspring, but did generate teratomas and differentiated embryoid bodies, and thus these specific iPSC lines were not completely reprogrammed or truly pluripotent. Therefore, there is a need to address whether the reprogramming factors and process used eventually to generate chimeric mice are universal and sufficient to generate reprogrammed iPSC that contribute to chimeric offspring in additional species. Here we show that porcine mesenchymal stem cells transduced with 6 human reprogramming factors (POU5F1, SOX2, NANOG, KLF4, LIN28, and C-MYC) injected into preimplantation-stage embryos contributed to multiple tissue types spanning all 3 germ layers in 8 of 10 fetuses. The chimerism rate was high, 85.3% or 29 of 34 live offspring were chimeras based on skin and tail biopsies harvested from 2- to 5-day-old pigs. The creation of pluripotent porcine iPSCs capable of generating chimeric offspring introduces numerous opportunities to study the facets significantly affecting cell therapies, genetic engineering, and other aspects of stem cell and developmental biology.
authors.affiliation: Regenerative Bioscience Center, University of Georgia, Athens, Georgia 30602, USA.
authors.forename: Franklin D
authors.initials: FD
authors.lastname: West
authors.forename: Steve L
authors.initials: SL
authors.lastname: Terlouw
authors.forename: Dae Jin
authors.initials: DJ
authors.lastname: Kwon
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Mumaw
authors.forename: Sujoy K
authors.initials: SK
authors.lastname: Dhara
authors.forename: Kowser
authors.initials: K
authors.lastname: Hasneen
authors.forename: John R
authors.initials: JR
authors.lastname: Dobrinsky
authors.forename: Steven L
authors.initials: SL
authors.lastname: Stice
mesh_terms: D000825:Animal Structures / Q000166:cytology / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn / Q000002:abnormalities / Q000378:metabolism; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics; D001755:Blastocyst / Q000166:cytology; D002454:Cell Differentiation / Q000235:genetics; D002678:Chimera / Q000002:abnormalities / Q000196:embryology* / Q000378:metabolism; D058732:Embryoid Bodies / Q000166:cytology; D005326:Fetal Proteins / Q000235:genetics; D005333:Fetus / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics; D059630:Mesenchymal Stem Cells / Q000166:cytology; D000071317:Nanog Homeobox Protein; D009419:Nerve Tissue Proteins / Q000235:genetics; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D034421:Sus scrofa; D020825:T-Box Domain Proteins / Q000235:genetics; D014161:Transduction, Genetic; D000509:alpha-Fetoproteins / Q000235:genetics; D000098908:Brachyury Protein
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D005326:Fetal Proteins; D018398:Homeodomain Proteins; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D000071317:Nanog Homeobox Protein; D009419:Nerve Tissue Proteins; D050814:Octamer Transcription Factor-3; D016271:Proto-Oncogene Proteins c-myc; D016601:RNA-Binding Proteins; D055748:SOXB1 Transcription Factors; D020825:T-Box Domain Proteins; D000509:alpha-Fetoproteins; D000098908:Brachyury Protein; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; C000715187:Lin28A protein, human; C489427:MYC protein, human; C486780:NANOG protein, human; C494173:POU5F1 protein, human; C527782:SOX2 protein, human; C093564:Neurogenic differentiation factor 1