title: Gnotobiotic Miniature Pig Interbreed Somatic Cell Nuclear Transfer for Xenotransplantation.
abstract: Transgenic animal producing technology has improved consistently over the last couple of decades. Among the available methods, somatic cell nuclear transfer (SCNT) technology was officially the most popular. However, SCNT has low efficiency and requires a highly skilled individual. Additionally, the allo-SCNT nuclear reprogramming mechanism is poorly understood in the gnotobiotic miniature pig, which is a candidate for xenotransplantation, making sampling in oocytes very difficult compared to commercial hybrid pigs. Therefore, interbreed SCNT (ibSCNT), which is a combination of miniature pig and commercial pig (Landrace based), was analyzed and was found to be similar to SCNT in terms of the rate of blastocyst formation (12.6% ± 2.9% vs. 15.5% ± 2.2%; p > 0.05). However, a significantly lower fusion rate was observed in the ibSCNT compared to normal SCNT with Landrace pig somatic cells (29.6% ± 0.8% vs. 65.0% ± 4.9%). Thus, the optimization of fusion parameters was necessary for efficient SCNT. Our results further revealed that ibSCNT by the whole-cell intracytoplasmic injection (WCICI) method had a significantly higher blastocyst forming efficiency than the electrofusion method (31.1 ± 8.5 vs. 15.5% ± 2.2%). The nuclear remodeling and the pattern of changes in acetylation at H3K9 residue were similar in both SCNT and ibSCNT embryos.
authors.affiliation: 1 Bio-Organ Research Center, Konkuk University , Seoul, Republic of Korea.|3 Jeonbuk Department of Inhalation Research, Korea Institute of Toxicology , Jeongeup, Republic of Korea.
authors.forename: Jeong Ho
authors.initials: JH
authors.lastname: Hwang
authors.affiliation: 2 Department of Animal Biotechnology, Konkuk University , Seoul, Republic of Korea.
authors.forename: Sang Eun
authors.initials: SE
authors.lastname: Kim
authors.affiliation: 4 Department of Biotechnology and Medical Engineering, National Institute of Technology , Rourkela, India .
authors.forename: Mukesh Kumar
authors.initials: MK
authors.lastname: Gupta
authors.affiliation: 1 Bio-Organ Research Center, Konkuk University , Seoul, Republic of Korea.|2 Department of Animal Biotechnology, Konkuk University , Seoul, Republic of Korea.
authors.forename: HoonTaek
authors.initials: H
authors.lastname: Lee
mesh_terms: D000818:Animals; D004624:Embryo Transfer; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology*; D005260:Female; D005856:Germ-Free Life; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D009865:Oocytes / Q000166:cytology / Q000502:physiology*; D013552:Swine / Q000145:classification / Q000196:embryology*; D013556:Swine, Miniature / Q000196:embryology*; D014183:Transplantation, Heterologous