Nuclear Transfer by Using a Laser-Assisted Zona Pellucida Piercing Technique in Mice

  • Kang, Ho-In (Department of Craniomaxillofacial Reconstructive Science Major, Dental Research Institute, and CLS21, Seoul National University School of Dentistry) ;
  • Choi, Young-Ju (Department of Craniomaxillofacial Reconstructive Science Major, Dental Research Institute, and CLS21, Seoul National University School of Dentistry) ;
  • Sung, Ji-Hye (Department of Craniomaxillofacial Reconstructive Science Major, Dental Research Institute, and CLS21, Seoul National University School of Dentistry) ;
  • Park, Sang-Kyu (Department of Craniomaxillofacial Reconstructive Science Major, Dental Research Institute, and CLS21, Seoul National University School of Dentistry) ;
  • Min, Byung-Moo (Department of Craniomaxillofacial Reconstructive Science Major, Dental Research Institute, and CLS21, Seoul National University School of Dentistry) ;
  • Roh, Sang-Ho (Department of Craniomaxillofacial Reconstructive Science Major, Dental Research Institute, and CLS21, Seoul National University School of Dentistry)
  • Published : 2007.09.30

Abstract

Somatic cells nuclear transfer (SCNT) is a useful tool in studies of developmental biology and animal cloning. However, SCNT experiments only are allowed to skilled technical experts. In this experiment, laser-assisted zona pellucida piercing tool (LASER) was applied in murine SCNT. LASER minimized the use of piezo-driven micromanipulator (PIEZO), reducing chances of problems caused by PIEZO pulses. LASER reduced time that took to pierce zona pellucida in removal of nucleus from oocyte and somatic cell injection, which might have taken longer time with PIEZO. Time and difficulties that took researcher of equivalent skilled for their experiments were decreased with LASER, and this might affect the improvement of embryonic development. (LASER, 6.2% versus PIEZO, 2.9%; P<0.05). Thus, these data support that the use of LASER can be used for zona pellucida piercing in murine SCNT program as an alternative to PIEZO.

Keywords

References

  1. Allegra, A., Monni, G., Zoppi, M.A., Curcio, P., Marino, A. and Volpes, A. Conjoined twins in a trichorionic quadruplet pregnancy after intracytoplasmic sperm injection and quarter laser-assisted zona thinning. Fertil Steril 87:189-193, 2007
  2. Anzai, M., Nishiwai, M., Yanagi, M., Nakashima, T., Kaneko, T., Taguchi, Y., Tokoro, M., Shin, S.W., Mitani, T., Kato, H., Matosumo, K., Nakagata N. and Iritani, A. Application of laser-assisted zona drilling to in vitro fertilization of cryopreserved mouse oocytes with spermatozoa from a sub fertile transgenic mouse. J Reprod Dev 52:601-606, 2006 https://doi.org/10.1262/jrd.18040
  3. Chatot, C.L., Ziomek, C.A., Bavister, B.D., Lewis, J.L. and Torres, I. An improved culture medium supports development of random-bred 1-cell mouse embryos in vitro. J Reprod FertiI 86:679-688, 1989 https://doi.org/10.1530/jrf.0.0860679
  4. Chen, S.-U., Chao, K.-H., Chang, C.-Y., Hsieh, F.-J., Ho, H.-N. and Yang, Y.-S. Technical aspects of the piezo, laser-assisted, and conventional methods for nuclear transfer of mouse oocytes and their efficiency and efficacy: piezo minimizes damage of the ooplasmic membrane at injection. J Exp Zool 301A:344-351, 2004 https://doi.org/10.1002/jez.a.20037
  5. Gabrielsen, A., Agerholm, I., Toft, B., HaId, F., Petersen, K., Aagaard, J., Feldinger, B., Lindenberg, S. and Fedder, J. Assisted hatching improves implantation rates on cryopreserved-thawed embryos. A randomized prospective study. Hum Reprod 19:2258-2262, 2004 https://doi.org/10.1093/humrep/deh434
  6. Kishigami, S., Bui H.T., Wakayama, S., Tokunaga, K., Thuan, N.B., Hikichi, T., Mizutani, E., Ohta, H., Sata, T. and Wakayama, T. Successful mouse cloning of an outbred strain by trichostatin A treatment after somatic nuclear transfer. J Reprod Dev 53:165-170, 2007 https://doi.org/10.1262/jrd.18098
  7. Lawitts, J.A. and Biggers, J.D. Optimization of mouse embryo culture media using simplex methods. J Reprod Fertil 91:543-556, 1991 https://doi.org/10.1530/jrf.0.0910543
  8. Rybouchkin, A., Heindryckx, B., Van der Elst, J. and Dhont, M. Developmental potential of cloned mouse embryos reconstructed by a conventional technique of nuclear injection. Reproduction 124:197-207, 2002 https://doi.org/10.1530/rep.0.1240197
  9. Tadir, Y. and Douglas-Hamilton D.H. Laser effects in the manipulation of human eggs and embryos for in vitro fertilization. Method in Cell BioI 82:409-431, 2007 https://doi.org/10.1016/S0091-679X(06)82014-5
  10. Wakayama, S., Ohta, H., Kishigami, S., Thuan, N.V., Hikichi, T., Mizutani, E., Miyake, M. and Wakayama, T. Establishment of male and female nuclear transfer embryonic stem cell lines from different mouse strains and tissues. Biol Reprod 72:932-936, 2005 https://doi.org/10.1095/biolreprod.104.035105
  11. Wakayama, T. and Yanagimachi, R. Effect of cytokinesis inhibitors, DMSO and the timing of oocyte activation on mouse cloning using cumulus cell nuclei. Reproduction 122:49-60, 2001 https://doi.org/10.1530/rep.0.1220049
  12. Wakayama, T. and Yanagimachi, R. Cloning the laboratory mouse. Cell Dev BioI 10:253-258, 1999 https://doi.org/10.1006/scdb.1998.0267
  13. Wakayama, T., Perry, A.C.F., Zuccotti, M., Johnsonk, K.R., Yanagimachi, R. Full-term development of mice from nucleated oocytes injected with cumulus cell nuclei. Nature 394:369-374, 1998 https://doi.org/10.1038/28615
  14. Zhou, Q., Boulanger, L. and Renard, J.P. A simplified method for the reconstruction of fully competent mouse zygotes from adult somatic donor nuclei. Cloning 2:35-44, 2000 https://doi.org/10.1089/15204550050145111