[Effects of cryoprotectants and sucrose dilution on the survival of unfertilized and fertilized mouse eggs after freezing and thawing].
We have studied the methods of cryopreservation and dilution of the unfertilized and fertilized eggs of ICR mouse. To improve the survival rate, we examined 8 types of methods with various combination of cryoprotectants, dilutions and cooling rates. The highest survival rate for unfertilized eggs was obtained when 1.5 M dimethylsulfoxide (DMSO) + 0.25 M sucrose was added to PBS as a cryoprotectant and when the eggs were diluted by adding PBS with 0.5 M sucrose at room temperature at 5-minute intervals in 5 steps. The total survival rate was 45.4% (p less than 0.01). In the case of fertilized eggs, the highest survival rate (72.6%) was obtained with 10% glycerol + 0.25 M sucrose as a cryoprotectant and a One-step dilution method. The most effective cooling rate was 0.3 degrees C per minute for either unfertilized or fertilized eggs.
Takada
H
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Department of Obstetrics and Gynecology, Asahikawa Medical College.
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D003451:Cryoprotective Agents / Q000008:administration & dosage / Q000494:pharmacology*; D005260:Female; D005615:Freezing; D051379:Mice; D008813:Mice, Inbred ICR; D009994:Osmolar Concentration; D010063:Ovum; D011309:Preservation, Biological / Q000379:methods*; D013395:Sucrose / Q000008:administration & dosage / Q000494:pharmacology*; D015053:Zygote
D003451:Cryoprotective Agents; D013395:Sucrose