Dinnyés A, Wallace G A, Rall W F
Agricultural Biotechnology Center, Gödöllö, Hungary.
Mol Reprod Dev. 1995 Apr;40(4):429-35. doi: 10.1002/mrd.1080400406.
We examined possible genotype effects on the survival of 8- to 16-cell mouse embryos isolated from four inbred strains (C57BL/6N, BALB/cAnN, DBA/2N, and C3H/HeN), a outbred stock (ICR), and various crosses after cryopreservation by vitrification or conventional slow freezing using glycerol solutions. The rates of in vitro development of C57BL/6N, BALB/cAnN, C3H/HeN, and ICR embryos to expanded blastocysts ranged from 86% to 94% after slow freezing and 85% to 97% after vitrification. The cryopreservation method did not significantly influence in vitro embryo survival after thawing (P > 0.05). Although genotype significantly influenced the in vitro survival of embryos (P = 0.008), this presumably resulted from an increased difficulty in assessing the quality grade of C3H/HeN embryos prior to cryopreservation. The rates in vivo development of C57BL/6N, BALB/cAnN, C3H/HeN, DBA/2N, and ICR embryos to normal day 18-19 fetuses ranged from 19% to 64% after slow freezing and from 18% to 63% after vitrification. The in vivo development of cryopreserved embryos was significantly influenced by cryopreservation method and genotype (P = 0.01 and P = 0.001, respectively). Vitrification yielded significantly higher rates of in vivo development than that after slow freezing (P > 0.05). In vivo development rates of DBA/2N and ICR female X B6D2F1 male embryos after cryopreservation were significantly higher than that of embryos from BALB/cAnN and C3H/HeN mice (P < 0.05). These results indicate that parental genotype exerts little or no effect on the ability of embryos to develop in vitro after vitrification or slow freezing. Differences in the ability of cryopreserved embryos to develop normally in vivo may reflect inherent genotype related differences in their post-implantation developmental potential and not their sensitivity to cryoinjury.
我们研究了基因型对从四个近交系（C57BL/6N、BALB/cAnN、DBA/2N和C3H/HeN）、一个远交群（ICR）以及各种杂交组合中分离出的8至16细胞期小鼠胚胎冷冻保存后的存活率的可能影响。冷冻保存采用玻璃化法或使用甘油溶液的传统慢速冷冻法。C57BL/6N、BALB/cAnN、C3H/HeN和ICR胚胎经慢速冷冻后发育至扩张囊胚的体外发育率为86%至94%，经玻璃化冷冻后为85%至97%。冷冻保存方法对解冻后胚胎的体外存活率没有显著影响（P>0.05）。尽管基因型对胚胎的体外存活率有显著影响（P = 0.008），但这可能是由于在冷冻保存前评估C3H/HeN胚胎质量等级的难度增加所致。C57BL/6N、BALB/cAnN、C3H/HeN、DBA/2N和ICR胚胎经慢速冷冻后发育至正常第18至19天胎儿的体内发育率为19%至64%，经玻璃化冷冻后为18%至63%。冷冻保存胚胎的体内发育受到冷冻保存方法和基因型的显著影响（分别为P = 0.01和P = 0.001）。玻璃化冷冻后的体内发育率显著高于慢速冷冻后的发育率（P>0.05）。DBA/2N和ICR雌性×B6D2F1雄性胚胎冷冻保存后的体内发育率显著高于BALB/cAnN和C3H/HeN小鼠的胚胎（P<0.05）。这些结果表明，亲代基因型对玻璃化或慢速冷冻后胚胎的体外发育能力影响很小或没有影响。冷冻保存胚胎在体内正常发育能力的差异可能反映了其植入后发育潜力中与基因型相关的固有差异，而不是它们对冷冻损伤的敏感性。
