Equilibrium vitrification of mouse embryos at various developmental stages.
Mol Reprod Dev
; 79(11): 785-94, 2012 Nov.
Article
in En
| MEDLINE
| ID: mdl-22987665
ABSTRACT
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.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Blastocyst
/
Cryopreservation
/
Embryo, Mammalian
/
Vitrification
/
Morula
Limits:
Animals
Language:
En
Journal:
Mol Reprod Dev
Journal subject:
BIOLOGIA MOLECULAR
/
MEDICINA REPRODUTIVA
Year:
2012
Document type:
Article
Affiliation country:
Japan