Cloning endangered gray wolves (Canis lupus) from somatic cells collected postmortem

• Published in: Theriogenology
• Main Authors: Oh, H.J., Kim, M.K., Jang, Goo, Kim, H.J., Hong, S.G., Park, J.E., Park, K., Park, C., Sohn, S.H., Kim, Dae-Yong, Shin, Nam Shik, Lee, Byeong Chun
• Other Authors: 이병천
• Format: Article in Journal/Newspaper
• Language: English
• Published: Elsevier 2008
• Subjects: Somatic cell nuclear transfer; Telomeric DNA; Canine oocyte; Gray wolf; Postmortem somatic cell; Canis lupus; gray wolf
• Online Access: https://hdl.handle.net/10371/8297; https://doi.org/10.1016/j.theriogenology.2008.04.032

The objective of the present study was to investigate whether nuclear transfer of postmortem wolf somatic cells into enucleated dog oocytes, is a feasible method to produce a cloned wolf. In vivo-matured oocytes (from domestic dogs) were enucleated and fused with somatic cells derived from culture of tissue obtained from a male gray wolf 6 h after death. The reconstructed embryos were activated and transferred into the oviducts of naturally synchronous domestic bitches. Overall, 372 reconstructed embryos were transferred to 17 recipient dogs; four recipients (23.5%) were confirmed pregnant (ultrasonographically) 23–25 d after embryo transfer. One recipient spontaneously delivered two dead pups and three recipients delivered, by cesarean section, four cloned wolf pups, weighing 450, 190, 300, and 490 g, respectively. The pup that weighed 190 g died within 12 h after birth. The six cloned wolf pups were genetically identical to the donor wolf, and their mitochondrial DNA originated from the oocyte donors. The three live wolf pups had a normal wolf karyotype (78, XY), and the amount of telomeric DNA, assessed by quantitative fluorescence in situ hybridization, was similar to, or lower than, that of the nuclear donor. In conclusion, the present study demonstrated the successful cloning of an endangered male gray wolf via interspecies transfer of somatic cells, isolated postmortem from a wolf, and transferred into enucleated dog oocytes. Therefore, somatic cell nuclear transfer has potential for preservation of canine species in extreme situations, including sudden death. This study was financially supported by KOSEF (grant #M10625030005-07N250300510). The authors are grateful for a graduate fellowship provided by the Korean MEST through the BK21 program We thank Seoul Grand Park Zoo for providing the donor cells and Y.H. Kim (NICEM) for assisting with microsatellite analysis.