Cloning missy: obtaining multiple offspring of a specific canine genotype by somatic cell nuclear transfer.
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.
Hossein
Mohammad Shamim
MS
SooAm Biotech Research Foundation, Gyeonggi-do, South Korea.
Jeong
Yeon Woo
YW
Park
Sun Woo
SW
Kim
Joung Joo
JJ
Lee
Eugine
E
Ko
Kyeong Hee
KH
Kim
Huen Suk
HS
Kim
Yeun Wook
YW
Hyun
Sang Hwan
SH
Shin
Taeyoung
T
Hawthorne
Lou
L
Hwang
Woo Suk
WS
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
D002118:Calcium