title: Putative embryonic stem cells derived from porcine cloned blastocysts using induced pluripotent stem cells as donors.
abstract: The establishment of porcine embryonic stem cells (ESCs) would have great impact in biomedical studies and preclinical trials through their use in genetic engineering. However, authentic porcine ESCs have not been established until now. In this study, a total of seven putative ESC lines were derived from porcine embryos of various origins, including in vitro fertilization, parthenogenetic activation, and, in particular, induced pluripotent stem (iPS) nuclear transfer (NT) from a donor cell with induced pluripotent stem cells (iPSCs). To characterize these cell lines, several assays including an assessment of intensive alkaline phosphatase activity, karyotyping, embryoid body formation, expression analysis of the pluripotency-associated markers, and the three germ layerassociated markers were performed. Based on quantitative polymerase chain reaction, the expression levels of REX1 and FGFR2 in iPS-NT lines were higher than those of cells of other origins. Additionally, only iPS-NT lines showed multiple aberrant patterns of nuclear foci elucidated by immunofluorescence staining of H3K27me3 as a marker of the state of X chromosome inactivation and a less mature form of mitochondria like naive ESCs, by transmission electron microscopy. Together, these data suggested that established putative porcine ESC lines generally exhibited a primed pluripotent state, like human ESCs. However, iPS-NT lines have especially unique characteristics distinct from other origins because they have more epigenetic instability and naive-like mitochondrial morphology than other putative ESC lines. This is the first study to establish and characterize the iPSC-derived putative ESC lines and compare them with other lines derived from different origins in pigs.
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, Republic of Korea.
authors.forename: Eunhye
authors.initials: E
authors.lastname: Kim
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, Republic of Korea.
authors.forename: Seon-Ung
authors.initials: SU
authors.lastname: Hwang
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, Republic of Korea.
authors.forename: Hyunju
authors.initials: H
authors.lastname: Yoo
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, Republic of Korea.
authors.forename: Junchul David
authors.initials: JD
authors.lastname: Yoon
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, Republic of Korea.
authors.forename: Yubyeol
authors.initials: Y
authors.lastname: Jeon
authors.affiliation: School of Life Sciences and Biotechnology, Korea University, Seoul, Republic of Korea.
authors.forename: Hyunggee
authors.initials: H
authors.lastname: Kim
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, Republic of Korea.
authors.forename: Eui-Bae
authors.initials: EB
authors.lastname: Jeung
authors.affiliation: Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute for Agriculture and Life Science, Seoul National University, Seoul, Republic of Korea. Electronic address: leeck@snu.ac.kr.
authors.forename: Chang-Kyu
authors.initials: CK
authors.lastname: Lee
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, Republic of Korea. Electronic address: shhyun@cbu.ac.kr.
authors.forename: Sang-Hwan
authors.initials: SH
authors.lastname: Hyun
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology* / Q000502:physiology; D049109:Cell Proliferation; D019976:Cloning, Organism; D053595:Embryonic Stem Cells / Q000502:physiology*; D061252:Feeder Cells; D018507:Gene Expression Regulation, Developmental; D057026:Induced Pluripotent Stem Cells / Q000502:physiology*; D007621:Karyotyping; D051379:Mice; D046529:Microscopy, Electron, Transmission; D008928:Mitochondria / Q000502:physiology; D010312:Parthenogenesis; D013552:Swine / Q000196:embryology*
keywords: Embryonic stem cell; Induced pluripotent stem cell; Pig; Pluripotency