Equilibrium vitrification of mouse embryos at various developmental stages
Article first published online: 15 OCT 2012
Copyright © 2012 Wiley Periodicals, Inc.
Molecular Reproduction and Development
Volume 79, Issue 11, pages 785–794, November 2012
How to Cite
Jin, B., Mochida, K., Ogura, A., Koshimoto, C., Matsukawa, K., Kasai, M. and Edashige, K. (2012), Equilibrium vitrification of mouse embryos at various developmental stages. Mol. Reprod. Dev., 79: 785–794. doi: 10.1002/mrd.22113
- Issue published online: 16 NOV 2012
- Article first published online: 15 OCT 2012
- Accepted manuscript online: 14 SEP 2012 02:11PM EST
- Manuscript Accepted: 7 SEP 2012
- Manuscript Received: 26 JUL 2012
- Grants-in-Aid for Scientific Research of Japan Society for the Promotion of Science
Previously, we developed a new method by which 2-cell mouse embryos can be vitrified in liquid nitrogen in a near-equilibrium state, and then kept at −80°C for several days. In the present study, we examined whether or not the method was effective for mouse embryos at other developmental stages. Eight-cell embryos, morulae, and expanded blastocysts of ICR mice were vitrified with ethylene glycol-based solutions, named EFSc because of their composition of ethylene glycol (30–40%, v/v) and FSc solution. The FSc solution was PB1 medium containing 30% (w/v) Ficoll PM-70 plus 1.5 M sucrose. The extent of equilibrium was assessed by examining how well vitrified embryos survived after being kept at −80°C. When 8-cell embryos and morulae were vitrified with EFS35c or EFS40c and then kept at −80°C, the survival rate was high even after 4 days in storage and remained high after re-cooling in liquid nitrogen. On the other hand, the survival of vitrified-expanded blastocysts kept at −80°C was low. Therefore, 8-cell embryos and morulae can be vitrified in a near-equilibrium state using the same method as for 2-cell embryos. A high proportion of C57BL/6J embryos at the 2-cell, 8-cell, and morula stages vitrified with EFS35c developed to term after transportation on dry ice, re-cooling in liquid nitrogen, and transfer to recipients. In conclusion, the near-equilibrium vitrification method, which is effective for 2-cell mouse embryos, is also effective for embryos at the 8-cell and morula stages. The method would enable handy transportation of vitrified embryos using dry ice. Mol. Reprod. Dev. 79: 785–794, 2012. © 2012 Wiley Periodicals, Inc.