title: Transgenesis and nuclear transfer using porcine embryonic germ cells.
abstract: Embryonic germ (EG) cells are undifferentiated stem cells isolated from cultured primordial germ cells (PGC). Porcine EG cell lines with capacities of both in vitro and in vivo differentiation have been established. Because EG cells can be cultured indefinitely in an undifferentiated state, they may be more suitable for nuclear donor cells in nuclear transfer (NT) than somatic cells that have limited lifespan in primary culture. Use of EG cells could be particularly advantageous to provide an inexhaustible source of transgenic cells for NT. In this study the efficiencies of transgenesis and NT using porcine fetal fibroblasts and EG cells were compared. The rate of development to the blastocyst stage was significantly higher in EG cell NT than somatic cell NT (94 of 518, 18.2% vs. 72 of 501, 14.4%). To investigate if EG cells can be used for transgenesis in pigs, green fluorescent protein (GFP) gene was introduced into porcine EG cells. Nuclear transfer embryos using transfected EG cells gave rise to blastocysts (29 of 137, 21.2%) expressing GFP based on observation under fluorescence microscope. The results obtained from the present study suggest that EG cell NT may have advantages over somatic cell NT, and transgenic pigs may be produced using EG cells.
authors.affiliation: Department of Physiology, Dankook University School of Medicine, Cheonan, Chungnam, Korea.
authors.forename: Kwang Sung
authors.initials: KS
authors.lastname: Ahn
authors.forename: Ji Young
authors.initials: JY
authors.lastname: Won
authors.forename: Soon Young
authors.initials: SY
authors.lastname: Heo
authors.forename: Jee Hyun
authors.initials: JH
authors.lastname: Kang
authors.forename: Hong Seok
authors.initials: HS
authors.lastname: Yang
authors.forename: Hosup
authors.initials: H
authors.lastname: Shim
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001755:Blastocyst; D018929:Cell Culture Techniques / Q000379:methods; D019976:Cloning, Organism / Q000379:methods; D004247:DNA / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology*; D018014:Gene Transfer Techniques; D005821:Genetic Techniques; D005854:Germ Cells / Q000166:cytology*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D053652:Nuclear Transfer Techniques; D013552:Swine; D019076:Transgenes
chemical_list: D049452:Green Fluorescent Proteins; D004247:DNA