title: Cloning missy: obtaining multiple offspring of a specific canine genotype by somatic cell nuclear transfer.
abstract: The present study was undertaken to evaluate two activation methods for somatic cell nuclear transfer (SCNT), namely, fusion and simultaneous activation (FSA, fusion medium contains calcium), versus fusion followed by chemical activation (F+CA, fusion medium does not contain calcium), and to evaluate the effects of parity of recipient dogs on the success of SCNT. Oocytes retrieved from outbred dogs were reconstructed with adult somatic cells collected from an 11-year-old female dog named Missy. In the FSA method, oocytes were fused and activated at the same time using two DC pulses of 1.75 kV/cm for 15 microsec. In the F+CA method, oocytes were fused with two DC pulses of 1.75 kV/cm for 15 microsec, and then activated 1 h after fusion by 10 microM calcium ionophore for 4 m and cultured for 4 h in 1.9 mM 6-dimethylaminopurine for postactivation. Activation method had a significant impact on the production efficiency of cloned dogs. There was a significant difference in full-term pregnancy rate and percentage of live puppies between the two methods (6.3% and 38.5% for FSA and F+CA, respectively). In our study, four out of five live offspring produced by F+CA survived versus FSA, which did not result in any surviving puppies. Overall, as few as 14 dogs and 54 reconstructed embryos were needed to produce a cloned puppy. In addition, the parity of recipient bitches had no effect on the success of SCNT in canine species. Both the nullipara and multipara bitches produced live puppies following SCNT-ET.
authors.affiliation: SooAm Biotech Research Foundation, Gyeonggi-do, South Korea.
authors.forename: Mohammad Shamim
authors.initials: MS
authors.lastname: Hossein
authors.forename: Yeon Woo
authors.initials: YW
authors.lastname: Jeong
authors.forename: Sun Woo
authors.initials: SW
authors.lastname: Park
authors.forename: Joung Joo
authors.initials: JJ
authors.lastname: Kim
authors.forename: Eugine
authors.initials: E
authors.lastname: Lee
authors.forename: Kyeong Hee
authors.initials: KH
authors.lastname: Ko
authors.forename: Huen Suk
authors.initials: HS
authors.lastname: Kim
authors.forename: Yeun Wook
authors.initials: YW
authors.lastname: Kim
authors.forename: Sang Hwan
authors.initials: SH
authors.lastname: Hyun
authors.forename: Taeyoung
authors.initials: T
authors.lastname: Shin
authors.forename: Lou
authors.initials: L
authors.lastname: Hawthorne
authors.forename: Woo Suk
authors.initials: WS
authors.lastname: Hwang
mesh_terms: D000818:Animals; D002118:Calcium / Q000737:chemistry; D019976:Cloning, Organism / Q000379:methods / Q000662:veterinary*; D004285:Dogs / Q000235:genetics*; D005838:Genotype; D018895:Microsatellite Repeats / Q000235:genetics; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D009865:Oocytes / Q000502:physiology
chemical_list: D002118:Calcium