Somatic Cell Nuclear Transfer of Oocytes Aspirated from Postovulatory Ovarian Follicles of Superovulated Rabbits

  • Shang, Jiang-Hua (Department of Animal Physiology, College of Biological Sciences, China Agricultural University) ;
  • Xu, Ru-Xiang (Neuromedical Institute, Zhujiang Hospital, Southern Medical University) ;
  • Jiang, Xiao-Dan (Neuromedical Institute, Zhujiang Hospital, Southern Medical University) ;
  • Zou, Yu-Xi (Neuromedical Institute, Zhujiang Hospital, Southern Medical University) ;
  • Qin, Ling-Sha (Neuromedical Institute, Zhujiang Hospital, Southern Medical University) ;
  • Cai, Ying-Qian (Neuromedical Institute, Zhujiang Hospital, Southern Medical University) ;
  • Yang, Zhi-Jun (Neuromedical Institute, Zhujiang Hospital, Southern Medical University) ;
  • Zheng, Xing (Department of Animal Physiology, College of Biological Sciences, China Agricultural University) ;
  • Cui, Sheng (Department of Animal Physiology, College of Biological Sciences, China Agricultural University)
  • 투고 : 2006.10.09
  • 심사 : 2006.12.17
  • 발행 : 2007.09.01


The aim of this study was to evaluate if oocytes, aspirated from postovulatory ovarian follicles of superovulated rabbits 14 h post-hCG administration, could be efficiently used as ooplasm recipients for somatic cell nuclear transfer (SCNT). Within a common SCNT protocol, a comparison between oocytes recovered by direct aspiration (aspirated) from available ovarian follicles and oocytes flushed out from oviducts (flushed) was carried out. The results showed that maturation and enucleation rates of aspirated oocytes were 70.7% and 69.2%, significantly lower than 95.3% (p<0.01) and 83.6% (p<0.05), respectively, from flushed oocytes. However, following enucleation of matured oocytes as ooplasm recipients for SCNT, no difference was recorded in fusion and cleavage rates, as well as blastocyst development from cleaved embryos or hatching of blastocysts between aspirated and flushed groups. Additionally, some matured aspirated and flushed oocytes were also used for immediate parthenogenetic activation and the resulting embryo development was not significantly different. Results from this study show the following: i) the majority of oocytes aspirated from postovulatory ovarian follicles of superovulated rabbits 14 h post-hCG administration are matured and can be used directly as ooplasm recipients for SCNT; ii) the reconstructed embryos derived from these oocytes have similar in vitro developmental ability to those flushed from the oviducts.


Nuclear Transfer;Oocyte;Rabbit;Follicle;Somatic Cell


연구 과제 주관 기관 : Natural Science Foundation of China, Natural Science Foundation for Outstanding Young Scientists of China


  1. Chen, D. Y., D. C. Wen, Y. P. Zhang, Q. Y. Sun, Z. M. Han, Z. H. Liu, P. Shi, J. S. Li, J. G. Xiangyu, L. Lian, Z. H. Kou, Y. Q. Wu, Y. C. Chen, P. Y. Wang and H. M. Zhang. 2002. Interspecies implantation and mitochondria fate of pandarabbit cloned embryos. Biol. Reprod. 67:637-642
  2. Campbell, K. H. S., R. Alberio, I. Choi, P. Fisher, R. D. W. Kelly, J. H. Lee and W. Maalouf. 2005. Cloning: eight years after Dolly. Reprod. Domest. Anim. 40:255-414
  3. Brackett, B. G., J. A. Mills and G. G. Jeitles. 1972. In vitro fertilization of rabbit ova recovered from ovarian follicles. Fertil. Steril. 23:898-909
  4. Bomsel-Helmreich, O., L. N. Huyen and I. Durand-Gasselin. 1989. Effects of varying doses of hCG on the evolution of preovulatory rabbit follicles and oocytes. Hum. Reprod. 4:636-642
  5. Lorenzo, P. L., P. G. Rebollar, M. J. Illera, J.C. Illera, M. Illera and J. M. Alvarino. 1996. Stimulatory effect of insulin-like growth factor I and epidermal growth factor on the maturation of rabbit oocytes in vitro. J. Reprod. Fertil. 107:109-117
  6. Kobayashi, Y., K. H. Wright, R. Santulli and E. E. Wallach. 1981. Ovulation and ovum maturation in the rabbit ovary perfused in vitro. Biol. Reprod. 24:483-490
  7. Lanza, R. P., J. B. Cibelli, F. Diaz, C. T. Moraes, P. W. Farin, C. E. Farin, C. J. Hammer, M. D. West and P. Damiani. 2000. Cloning of an endangred species (Bos gaurus) using interspecies nuclear transfer. Cloning 2:79-90
  8. Jiang, Y., T. Chen, C. L. Nan, Y. C. Ouyang, Q. Y. Sun and D. Y. Chen. 2005. In vitro culture and mtDNA fate of ibex-rabbit nuclear transfer embryos. Zygote. 13:233-240
  9. Hong, S. B., S. J. Uhm, H. Y. Lee, C. Y. Park, M. K. Gupta, B. H. Chung, K. S. Chung and H. T. Lee. 2005. Developmental ability of bovine embryos nuclear transferred with frozenthawed or cooled donor cells. Asian-Aust. J. Anim. Sci. 18:1242-1248
  10. Hewitson, L., T. Dominko, D. Takahashi, C. Martinovich, J. Ramalho-Santos, P. Sutovsky, J. Fanton, D. Jacob, D. Monteith, M. Neuringer, D. Battaglia, C. Simerly and G. Schatten. 1999. Unique checkpoints during the first cell cycle of fertilization after intracytoplasmic sperm injection in rhesus monkeys. Nat. Med. 5:431-433
  11. Hayashi, S., Y. Noda, H. Matsumoto and T. Mori. 1987. Fertilizability of unovulated mature eggs following indomethacin administration in mice. Gamete Res. 18:291-299
  12. Hardarson, T., K. Lundin and L. Hamberfer. 2000. The position of the metaphase II spindle cannot be predicted by the location of the first polar body in the human oocyte. Hum. Reprod. 15:1372-1376
  13. Harper, M. J. K. 1963. Ovulation in rabbits: the time of follicular rupture and expulsion of the eggs, in relation to injection of luteinizing hormone. J. Endocrinol. 26:307-316
  14. Dinnyes, A., Y. P. Dai, M. Barber, L. Liu, J. Xu, P. L. Zhou and X. Z. Yang. 2001. Development of cloned embryos from adult rabbit fibroblasts effect of activation treatment and donor cell preparation. Biol. Reprod. 64:257-263
  15. Moon, J. H., B. C. Jee, S. Y. Ku, C. S. Suh, S. H. Kim, Y. M. Choi, J. G. Kim and S. Y. Moon. 2005. Spindle positions and their distributions in in vivo and in vitro matured mouse oocytes. Hum. Reprod. 20:2207-2210
  16. Mitalipov, S. M., K. L. White, V. R. Farrar, J. Morrey and W. A. Reed. 1999. Development of nuclear transfer and parthenogenetic rabbit embryos activated with inositol 1,4,5- trisphosphate. Biol. Reprod. 60:821-827
  17. Rienzi, L., F. Martinez, F. Ubaldi, F. Martinea, M. Iacobelli, M. G. Minasi, S. Ferrero, J. Tesarik and E. Greco. 2003. Relationship between meiotic spindle location with regard to the polar body position and oocyte developmental potential after ICSI. Hum. Reprod. 18:1289-1293
  18. Sun, Q. Y., L. Lai, K. W. Park, B. Kuhholzer, R. S. Prather and H. Schatten. 2001. Dynamic events are differently mediated by microfilaments, microtubules, and mitogen-activated protein kinase during porcine oocyte maturation and fertilization in vitro. Biol. Reprod. 64:879-889
  19. Silva, C. P., K. Kommineni, R. Oldenbourg and D. L. Keefe. 1999. The first polar body does not predict accurately the location of the metaphase II meiotic spindle in mammalian oocytes. Fertil. Steril. 71:719-721
  20. Varian, N. B., R. R. Maurer and R. H. Foote. 1967. Ovarian response and cleavage rate of ova in control and FSH-primed rabbits receiving varying levels of luteinizing hormone. J. Reprod. Fertil. 13:67-73
  21. Wakayama, T., V. Tabar, I. Rodriguez, A. C. Perry, L. Studer and P. Mombaerts. 2001. Differentiation of embryonic stem cell lines generated from adult somatic cells by nuclear transfer. Sci. 292:740-743
  22. Yin, X. J., Y. Kato and Y. Tsunoda. 2001. Effect of enucleation procedures and maturation conditions on the development of nuclear-transferred rabbit oocytes receiving male fibroblast cell. Reprod. 124:41-47
  23. Chen, Y., Z. X. He, A. Liu, K. Wang, W. W. Mao, J. X. Chu, Y. Lu, Z. F. Fang, Y. T. Shi and Q. Z. Yang. 2003. Embryonic stem cells generated by nuclear transfer of human somatic nuclei into rabbit oocytes. Cell Res. 13:251-264
  24. Brophy, B., G. Smolenski, T. Wheeler, D. Wells, P. L'Huillier and G. Laible. 2003. Cloned transgenic cattle produce milk with higher levels of beta-casein and kappa-casein. Nat. Biotechnol. 21:157-162
  25. Oh, Y. K. and B. G. Brackett. 1975. Ultrastructure of rabbit ova recovered from ovarian follicles and inseminated in vitro. Fertil. Steril. 26:665-85
  26. Yang, C. X., Z. M. Han, D. C. Wen, Q. Y. Sun, K. Y. Zhang, L. S. Zhang, Y. Q. Wu, Z. H. Kou and D. Y. Chen. 2003. In vitro development and mitochondrial fate of macaca-rabbit cloned embryos. Mol. Reprod. Dev. 65:396-401