title: Epigenetic changes of lentiviral transgenes in porcine stem cells derived from embryonic origin.
abstract: Because of the physiological and immunological similarities that exist between pigs and humans, porcine pluripotent cell lines have been identified as important candidates for preliminary studies on human disease as well as a source for generating transgenic animals. Therefore, the establishment and characterization of porcine embryonic stem cells (pESCs), along with the generation of stable transgenic cell lines, is essential. In this study, we attempted to efficiently introduce transgenes into Epiblast stem cell (EpiSC)-like pESCs. Consequently, a pluripotent cell line could be derived from a porcine-hatched blastocyst. Enhanced green fluorescent protein (EGFP) was successfully introduced into the cells via lentiviral vectors under various multiplicities of infection, with pluripotency and differentiation potential unaffected after transfection. However, EGFP expression gradually declined during extended culture. This silencing effect was recovered by in vitro differentiation and treatment with 5-azadeoxycytidine. This phenomenon was related to DNA methylation as determined by bisulfite sequencing. In conclusion, we were able to successfully derive EpiSC-like pESCs and introduce transgenes into these cells using lentiviral vectors. This cell line could potentially be used as a donor cell source for transgenic pigs and may be a useful tool for studies involving EpiSC-like pESCs as well as aid in the understanding of the epigenetic regulation of transgenes.
authors: Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute for Agriculture and Life Science, Seoul National University, Seoul, Korea.
authors: Kwang-Hwan
authors: KH
authors: Choi
authors: Jin-Kyu
authors: JK
authors: Park
authors: Hye-Sun
authors: HS
authors: Kim
authors: Kyung-Jun
authors: KJ
authors: Uh
authors: Dong-Chan
authors: DC
authors: Son
authors: Chang-Kyu
authors: CK
authors: Lee
mesh_terms: D000818:Animals; D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D015415:Biomarkers / Q000378:metabolism; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D019175:DNA Methylation; D000077209:Decitabine; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D044127:Epigenesis, Genetic; D061252:Feeder Cells; D005822:Genetic Vectors; D005855:Germ Layers / Q000166:cytology / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D013552:Swine; D019076:Transgenes
chemical_list: D015415:Biomarkers; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D000077209:Decitabine; D001374:Azacitidine
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