title: Porcine Pluripotent Stem Cells Derived from IVF Embryos Contribute to Chimeric Development In Vivo.
abstract: Although the pig is considered an important model of human disease and an ideal animal for the preclinical testing of cell transplantation, the utility of this model has been hampered by a lack of genuine porcine embryonic stem cells. Here, we derived a porcine pluripotent stem cell (pPSC) line from day 5.5 blastocysts in a newly developed culture system based on MXV medium and a 5% oxygen atmosphere. The pPSCs had been passaged more than 75 times over two years, and the morphology of the colony was similar to that of human embryonic stem cells. Characterization and assessment showed that the pPSCs were alkaline phosphatase (AKP) positive, possessed normal karyotypes and expressed classic pluripotent markers, including OCT4, SOX2 and NANOG. In vitro differentiation through embryonic body formation and in vivo differentiation via teratoma formation in nude mice demonstrated that the pPSCs could differentiate into cells of the three germ layers. The pPSCs transfected with fuw-DsRed (pPSC-FDs) could be passaged with a stable expression of both DsRed and pluripotent markers. Notably, when pPSC-FDs were used as donor cells for somatic nuclear transfer, 11.52% of the reconstructed embryos developed into blastocysts, which was not significantly different from that of the reconstructed embryos derived from porcine embryonic fibroblasts. When pPSC-FDs were injected into day 4.5 blastocysts, they became involved in the in vitro embryonic development and contributed to the viscera of foetuses at day 50 of pregnancy as well as the developed placenta after the chimeric blastocysts were transferred into recipients. These findings indicated that the pPSCs were porcine pluripotent cells; that this would be a useful cell line for porcine genetic engineering and a valuable cell line for clarifying the molecular mechanism of pluripotency regulation in pigs.
authors.affiliation: College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
authors.forename: Binghua
authors.initials: B
authors.lastname: Xue
authors.affiliation: College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
authors.forename: Yilong
authors.initials: Y
authors.lastname: He
authors.affiliation: College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
authors.forename: Renyue
authors.initials: R
authors.lastname: Wei
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, 150081, China.
authors.forename: Ruizhen
authors.initials: R
authors.lastname: Sun
authors.affiliation: College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Yin
authors.affiliation: College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
authors.forename: Gerelchimeg
authors.initials: G
authors.lastname: Bou
authors.affiliation: College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Liu
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D002678:Chimera; D004622:Embryo, Mammalian / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D005347:Fibroblasts; D005855:Germ Layers / Q000166:cytology; D051379:Mice; D008819:Mice, Nude; D053652:Nuclear Transfer Techniques / Q000662:veterinary; D039904:Pluripotent Stem Cells / Q000166:cytology; D011247:Pregnancy; D013552:Swine / Q000196:embryology*; D013724:Teratoma