两种玻璃化液冻存小鼠桑椹胚的研究

目的:研究EDS-40和EFS-40两种玻璃化溶液对小鼠桑椹胚冻存的影响。方法:随机选用NIH小鼠优质桑椹胚,先对两种玻璃化溶液EDS-40(40%乙二醇+18%葡聚糖+0.5mol蔗糖)和EFS-40(40%乙二醇+18%聚蔗糖+0.5mol蔗糖)行毒性测试:再在20℃±5℃室温下,将小鼠桑椹胚分别加入到两种玻璃化溶液,玻璃化后装管并迅速投入液氮中,冻存2-3个月。各组均在25℃的水浴中复温,胚胎用解冻液(S-PBS)和培养液反复洗涤后移入含Ham'sF12培养液培养48h,观察胚胎形态及发育。部分胚胎体外培养12-14h后行胚胎移植,观察妊娠及产仔情况。结果:两种玻璃化溶液的毒性有显著性差异,EDS-40较EFS-40毒性低(X2=6.415,P<0.05),透明带完好率两组间无显著性差异(X2=2.189,P>0.05)。胚胎优良率(X2=7.51,P<0.01)和发育率(X2=5.556,P<0.05)EDS-40较EFS-40更好,两组间妊娠率(X2=1.937,P>0.05)及产仔率(X2=0.245,P>0.05)无显著性差异。结论:EDS-40较EFS-40玻璃化溶液毒性低,冻存胚胎效果更好。

Comparison of Efficacy Between Two Vitrification Solutions for Mouse Morulae

Objective: To study the efficacy of two vitrification solutions for mouse morulae. Methods: Good morulaes of NM mice were collected and used to test toxicity of the vitrification solutions EDS-40 (40% ethylene glycol,18% dextran and 0.5 mol sucrose) and EFS-40 (40% ethylene glycol,18% ficoll and 0.5 mol sucrose). Fine vitrified morulae were packaged in 0.25 mL plastic straws and immersed into liquid nitrogen and cryopreserved for about 2-3 months. Then the straws were heated rapidly, washed in Ham's F12 medium and cultured. The viability was determined by morphology and blastocyst formation after being cultured for 48 h. Some embryos were transplanted to recipients after being cultured for 12-14 h. The number of pregnant recipients and young born was counted and analyzed by Chi-squared test. Results: The toxicity of EDS-40 solution was significantly lower than that of EFS-40 (P<0.05) and the number of embryos developed to the blastocysts after vitrification in EDS-40 was significantly higher than in EFS-40 (P<0.05). The number of zona pellucida and the number of pregnancy and birth integrated after vitrification cryopreservation had no significant difference between EDS-40 and EFS-40 (P>0.05). However, the embryo fineness rates after vitrification in EDS-40 was significantly better than in EFS-40 (P<0.01). Conclusion: EDS-40 solution has less toxicity and better cryoprotect effect on embryos than EFS-40.