title: Treatment of fetal fibroblasts with DNA methylation inhibitors and/or histone deacetylase inhibitors improves the development of porcine nuclear transfer-derived embryos.
abstract: This study investigated whether treating fetal fibroblast cells (donor cells) with epigenetic modification-inducing drugs could improve the development of porcine cloned embryos. Donor cells were treated with different DNA methylation inhibitors (5-aza-dC, zebularine or RG108; 5nM) or histone deacetylase inhibitors (TSA, NaBu or SCR; 50nM) for 1h, and then subjected to SCNT. All of the treated groups showed significantly higher blastocyst formation rates compared to the control group. We chose 5-aza-dC and TSA as a combined treatment, and found that donor cells co-treated with 2.5nM 5-aza-dC for 1h and subsequently treated with 50nM TSA for another 1h before SCNT showed significantly improved blastocyst rates compared to the control, 5-aza-dC-treated, and TSA-treated groups. The levels of DNA methylation were decreased (though not to a significant degree) in donor cells treated with 5-aza-dC, TSA or both. The histone H3 acetylation levels were significantly increased in donor cells treated with TSA or co-treated with 5-aza-dC and TSA. Donor cells simultaneously co-treated with 5nM 5-aza-dC and 50nM TSA for 1h showed increased apoptosis of SCNT blastocysts. However, when we decreased the concentration of 5-aza-dC to 2.5nM, the co-treatment induced less apoptosis among SCNT blastocysts and the blastocyst development rate improved. Together, these results indicate that treatment of donor cells with 5-aza-dC, TSA, or TSA plus a low dose of 5-aza-dC could improve the blastocyst development of porcine cloned embryos.
authors.affiliation: Department of Animal Science & Biotechnology, Chungnam National University, Daejeon City 338-708, Republic of Korea.
authors.forename: Yun-Fei
authors.initials: YF
authors.lastname: Diao
authors.forename: Ken-Ji
authors.initials: KJ
authors.lastname: Naruse
authors.forename: Rong-Xun
authors.initials: RX
authors.lastname: Han
authors.forename: Xiao-Xia
authors.initials: XX
authors.lastname: Li
authors.forename: Reza K
authors.initials: RK
authors.lastname: Oqani
authors.forename: Tao
authors.initials: T
authors.lastname: Lin
authors.forename: Dong-Il
authors.initials: DI
authors.lastname: Jin
mesh_terms: D000818:Animals; D019976:Cloning, Organism; D003562:Cytidine / Q000031:analogs & derivatives / Q000494:pharmacology; D019175:DNA Methylation / Q000187:drug effects*; D046149:Embryo Culture Techniques / Q000662:veterinary*; D047108:Embryonic Development / Q000187:drug effects; D057890:Epigenomics; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D010797:Phthalimides / Q000494:pharmacology; D013552:Swine / Q000196:embryology*; D014364:Tryptophan / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D056572:Histone Deacetylase Inhibitors; D010797:Phthalimides; C503639:RG108; D003562:Cytidine; C009131:pyrimidin-2-one beta-ribofuranoside; D014364:Tryptophan
keywords: 5-aza-dC; Embryo development; Epigenetic modification; Porcine nuclear transfer; TSA