title: Combined Treatment with Demecolcine and 6-Dimethylaminopurine during Postactivation Improves Developmental Competence of Somatic Cell Nuclear Transfer Embryos in Pigs.
abstract: This study determined the effects of postactivation treatment with demecolcine and/or 6-dimethylaminopurine (6-DMAP) on in vivo and in vitro developmental competence of somatic cell nuclear transfer (SCNT) embryos in pigs. SCNT embryos were treated for 4 hours with 0.4 µg/mL demecolcine, 2 mM 6-DMAP, or both after electric activation, then transferred to surrogate pigs or cultured for 7 days. The formation rate of SCNT embryos with a single pronucleus was higher in combined treatment with demecolcine and 6-DMAP (95.2%) than treatment with demecolcine alone (87.1%). Blastocyst formation of SCNT embryos was significantly increased in combined treatment with demecolcine and 6-DMAP (48.7%) compared with demecolcine (22.2%) or 6-DMAP alone (37.3%). Fluctuation of maturation promoting factor activity showed different patterns among various postactivation treatments. Pregnancy was established in 1 of 5 surrogates after transfer of SCNT embryos that were treated with demecolcine and 6-DMAP. The pregnant surrogate delivered one healthy live piglet. The results of our study demonstrated that postactivation treatment with demecolcine and 6-DMAP together improved preimplantation development and supported normal in vivo development of SCNT pig embryos, probably influencing MPF activity and nuclear remodeling, including induction of single pronucleus formation after electric activation.
authors.affiliation: a College of Veterinary Medicine , Kangwon National University , Chuncheon , Korea.|b Institute of Veterinary Science , Kangwon National University , Chuncheon , Korea.
authors.forename: Joohyeong
authors.initials: J
authors.lastname: Lee
authors.affiliation: a College of Veterinary Medicine , Kangwon National University , Chuncheon , Korea.
authors.forename: Jinyoung
authors.initials: J
authors.lastname: You
authors.affiliation: a College of Veterinary Medicine , Kangwon National University , Chuncheon , Korea.
authors.forename: Geun-Shik
authors.initials: GS
authors.lastname: Lee
authors.affiliation: c College of Animal Life Science , Kangwon National University , Chuncheon , Korea.
authors.forename: Seung Tae
authors.initials: ST
authors.lastname: Lee
authors.affiliation: d College of Veterinary Medicine , Chungbuk National University , Cheongju , Korea.
authors.forename: Sang-Hwan
authors.initials: SH
authors.lastname: Hyun
authors.affiliation: a College of Veterinary Medicine , Kangwon National University , Chuncheon , Korea.|b Institute of Veterinary Science , Kangwon National University , Chuncheon , Korea.
authors.forename: Eunsong
authors.initials: E
authors.lastname: Lee
mesh_terms: D000225:Adenine / Q000008:administration & dosage / Q000031:analogs & derivatives*; D000818:Animals; D002467:Cell Nucleus / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D003703:Demecolcine / Q000008:administration & dosage*; D004624:Embryo Transfer / Q000379:methods / Q000662:veterinary*; D047108:Embryonic Development / Q000187:drug effects* / Q000502:physiology; D005260:Female; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D013552:Swine; D016896:Treatment Outcome; D050257:Tubulin Modulators / Q000008:administration & dosage
chemical_list: D050257:Tubulin Modulators; C001020:N(6),N(6)-dimethyladenine; D000225:Adenine; D003703:Demecolcine
keywords: Maturation promoting factor; nuclear remodeling; postactivation; somatic cell nuclear transfer