猪MⅡ期卵母细胞玻璃化冷冻的研究

Studies on MⅡStage Porcine Oocytes Vitrification

本研究旨在为猪MⅡ期卵母细胞的玻璃化冷冻选择合适的冷冻保护剂及冷冻载体,提高猪卵母细胞冷冻后的存活力。试验分别采用以40%EG、15%EG+15%DMSO+20%FBS、15%EG+15%DMSO+20%SPS作为渗透性冷冻保护剂的冷冻液,以玻璃微细管(GMP)和冷冻帽(cryotop)作为载体对猪卵母细胞进行玻璃化冷冻。结果发现,采用含有血清蛋白替代物(serum protein substitute,SPS)的冷冻液VS3冷冻后二乙酰荧光素(FDA)染色存活率(59.40%±4.93%)显著高于VS1组(42.12%±4.08%)及VS2组(37.57%±1.21%)(P<0.05);采用VS3作为冷冻液用cryotop作为载体冷冻后FDA染色存活率(83.33%±3.33%)显著高于GMP(59.40%±4.93%)(P<0.05)。由此可见,采用SPS代替胎牛血清(FBS),用cryotop作为载体可以使猪卵母细胞冻后存活率显著提高。

The study was designed to select right cryoprotectants and carrier for porcine matured oocytes to improve oocyte viability after vitrification and lay a good foundation for porcine oocytes continue development. Using 40 % EG, 15 % EG＋ 15 % DMSO＋20% FBS, 15% EG＋15% DMSO＋20% SPS as cryoprotectants and taking GMP and cryotop as carrier to do porcine oocytes vitrification. The results showed that VS3 including SPS made better active of FDA （59.40% ± 4.93%） than VS1 （42.12% ±4.08%） and VS2 （37.57 % ±1.21% ）（P〈0. 05）;VS3 as cryoprotectants and cryotop as carrier made better active of FDA （83.33%±3.33%） than GMP （59.40%±4. 93%）（P〈0.05）. The data demonstrated that use SPS to instead of FBS and cryotop as carrier could significantly improve active of oocyte after vitrification.