In vitro viability of mouse zygotes vitrified in ethylene glycol.
A study was made to determine if mouse zygotes can be effectively vitrified in 7 M ethylene glycol in modified Dulbecco's phosphate buffered saline (PB1) and to find out if the development of vitrified-warmed zygotes in vitro can be improved by renewing the culture medium. The results showed that without medium change, vitrification reduced the development of zygotes to the expanded blastocyst stage (p < 0.01). With medium change, the development rate of vitrified-warmed zygotes exposed in 7 M ethylene glycol for 1 or 2 min was similar to that of unvitrified zygotes. However, prolonged exposure (5 min) markedly reduced the development rates of vitrified-warmed zygotes to the expanded blastocyst stage (p < 0.05). When the zygotes were vitrified in 7 M ethylene glycol and diluted at 18 degrees C to 22 degrees C, a slower efflux of ethylene glycol from the cell might have occurred, leading to a toxic effect of ethylene glycol in culture. The development rates of vitrified embryos cultured with medium change at 24 hr did not significantly differ from the untreated control (89.0% vs 96.5%). In conclusion, this study showed that mouse zygotes can be vitrified in 7 M ethylene glycol in PB1 and that changing the culture medium can improve the in vitro development rates of vitrified-warmed zygotes to the expanded blastocyst stage.
Bautista
J A
JA
Department of Veterinary, Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan. jobau@vetmed.hokudai.ac.jp
Takahashi
Y
Y
Kanagawa
H
H
D000818:Animals; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D003470:Culture Media; D019855:Ethylene Glycol / Q000494:pharmacology*; D005260:Female; D051379:Mice; D008810:Mice, Inbred C57BL / Q000196:embryology*; D013997:Time Factors; D014783:Viscosity; D015053:Zygote / Q000187:drug effects / Q000254:growth & development*
D003451:Cryoprotective Agents; D003470:Culture Media; D019855:Ethylene Glycol