摘要
本试验用10%或20%GL预处理5min,再移入GFS40中平衡0.5min二步法冷冻保存的小鼠孵化囊胚,解冻后24h内恢复率高达93%—97%,与对照组(99%)相比无显著差异(P>0.05)。新鲜孵化囊胚在低渗液中平衡30min,0.25×PBS组恢复率达92%,而0.20×PBS组恢复率仅为50%,与对照组相比差异极显著(P<0.01)。冷冻解冻后孵化囊胚直接进行低渗处理,结果0.50×PBS组的恢复率(72%)与对照组(88%)差异显著(P<0.05),而0.30×、0.25×和0.20×PBS组的恢复率分别为58%、11%和0%。解冻后经培养的孵化囊胚对低渗处理的耐受力增强,0.50×PBS处理组的恢复率达94%,与对照组(98%)相比无显著差异(P>0.05),0.30×、0.25×和0.20×PBS组的恢复率与对照组相比虽有极显著差异(P<0.01),但与解冻后直接进行低渗处理组相比恢复率有较大提高(82%—26%)。
The objective of this study to evaluate the effect of hypotonic stress on developmental potential of hatched blastocysts perivitrification. Hatched mouse blastocysts were vitrified in liquid nitrogen after equilibration in 10% or 20% GL for 5min and in GFS40 for 30sec respectively, the survival rates were 93%- 97% after the frozen-thawed embryos were cultured in vitro for 24h. There were no statistical difference between the frozen and the fresh group (P>0. 05). In order to evaluate effects of hypotonic stress on developmental abilities, fresh hatched mouse blastocysts were respectively exposed to 1. 00×0. 50×, 0. 30×, 0. 25×and 0. 20×PBS for 30min, then cultured in mKRB for 24h, the survival rates were 98%, 99%, 92%, 92% and 50% respectively. The rate in 0. 20X PBS group was significantly lower than in other groups (P<0. 01).
When frozen-thawed embryos were directly treated with different osmotic solutions, the survival rates were 88%, 72%, 58%, 11% and 0 respectively in 1. 00×, 0.50×, 0. 30×, 0. 25×and 0. 20×PBS group. The rate in 1.00×PBS group was significantly higher than in other groups (P < 0.05). However, when frozen-thawed embryos were first cultured in vitro for 12h, then exposed to 1. 00×, 0. 50×, 0. 30×, 0. 25×and 0. 2×PBS, the survival rates were 98%, 94%, 82%, 58% and 26% respectively. There was no statistical difference between 1. 00×and 0.50×PBS group (P>0. 05). Although the rate in 0. 30 ×, 0. 25×and 0. 20×PBS group was significantly lower than in 1.00× group(P<C 0. 01 ), it was significantly higher than in the same treatment group without in vitro culture (P <0. 05).
出处
《实验生物学报》
CSCD
2000年第3期215-221,共7页
Acta Biologiae Experimentalis Sinica
基金
国家"九五"攻关项目
合同号96-A23-06-05
