title: Leukemia inhibitory factor (LIF)-dependent, pluripotent stem cells established from inner cell mass of porcine embryos.
abstract: The pig is important for agriculture and as an animal model in human and veterinary medicine, yet despite over 20 years of effort, there has been a failure to generate pluripotent stem cells analogous to those derived from mouse embryos. Here we report the production of leukemia inhibitory factor-dependent, so-called naive type, pluripotent stem cells from the inner cell mass of porcine blastocysts by up-regulating expression of KLF4 and POU5F1. The alkaline phosphatase-positive colonies resulting from reprogramming resemble mouse embryonic stem cells in colony morphology, cell cycle interval, transcriptome profile, and expression of pluripotent markers, such as POU5F1, SOX2, and surface marker SSEA1. They are dependent on leukemia inhibitory factor signaling for maintenance of pluripotency, can be cultured over extended passage, and have the ability to form teratomas. These cells derived from the inner cell mass of pig blastocysts are clearly distinct from the FGF2-dependent "primed" induced pluripotent stem cells described recently from porcine mesenchymal cells. The data are consistent with the hypothesis that the up-regulation of KLF4, as well as POU5F1, is required to create and stabilize the naive pluripotent state and may explain why the derivation of embryonic stem cells from pigs and other ungulates has proved so difficult.
authors.affiliation: Division of Animal Sciences, University of Missouri, Columbia, Missouri 65211-7310. Electronic address: telugub@missouri.edu.
authors.forename: Bhanu Prakash V L
authors.initials: BPVL
authors.lastname: Telugu
authors.affiliation: Division of Animal Sciences, University of Missouri, Columbia, Missouri 65211-7310.
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Ezashi
authors.affiliation: Division of Animal Sciences, University of Missouri, Columbia, Missouri 65211-7310.
authors.forename: Sunilima
authors.initials: S
authors.lastname: Sinha
authors.affiliation: Division of Animal Sciences, University of Missouri, Columbia, Missouri 65211-7310.
authors.forename: Andrei P
authors.initials: AP
authors.lastname: Alexenko
authors.affiliation: Division of Animal Sciences, University of Missouri, Columbia, Missouri 65211-7310.
authors.forename: Lee
authors.initials: L
authors.lastname: Spate
authors.affiliation: Division of Animal Sciences, University of Missouri, Columbia, Missouri 65211-7310.
authors.forename: Randall S
authors.initials: RS
authors.lastname: Prather
authors.affiliation: Division of Animal Sciences, University of Missouri, Columbia, Missouri 65211-7310; Department of Biochemistry, University of Missouri, Columbia, Missouri 65211-7310. Electronic address: robertsrm@missouri.edu.
authors.forename: R Michael
authors.initials: RM
authors.lastname: Roberts
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism; D001755:Blastocyst / Q000166:cytology* / Q000378:metabolism; D054337:Cell Dedifferentiation / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D006801:Humans; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000378:metabolism; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology*; D008648:Mesoderm / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D013045:Species Specificity; D013552:Swine
chemical_list: D000943:Antigens, Differentiation; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D053605:Leukemia Inhibitory Factor; D050814:Octamer Transcription Factor-3