精子冷冻环无保护剂玻璃化冷冻方法的初步研究

Pilot study of cryoloop for vitrifying spermatozoa without cryoprotectants

目的探讨精子冷冻环无保护剂玻璃化冷冻方法的可行性。方法正常精液标本上游处理后,进行常规冷冻和冷冻环无保护剂的玻璃化冷冻,复苏后分别从活力参数及电镜下超微结构等指标比较两种冷冻方法的效果。结果两种方法冷冻后精子存活率、活动率之间差异无显著性(48%∶48%;44.5%∶43.5%,P>0.05),但均较未冷冻的89%和88.5%明显下降(P<0.001)。超微结构亦较未冷冻时发生了一定的改变,但核结构基本保持完整。结论精子冷冻环无保护剂的玻璃化冷冻是一种简单、方便而行之有效的冷冻方法。

Objective： To explore the feasibility of cryoloop for vitrifying spermatozoa without eryoproteetants, and seek for a suitable freezing carrier for testieular or epididymal sperm. Methods： Normal sperm were frozen by conventional freezing protocol and vitrification without cryoprotectants respectively. The mobility, viability and ultrastructures of frozenthawed sperm of two freezing protocols were compared. Results： Motility and viability were of no statistic differences between the two freezing groups （48% vs 48% and 44.5% vs 43.5%, PS〉0.05）, but lower evidently than those of the control group （48% vs 48% vs 89% and 44.5% vs 43.5% vs 88.5%, P〈0. 001）. The ultrastructures of frozen-thawed samples also changed, but the structure of nucleus remained intact. Conclusions： Non-cryoprotectant vitrification with cryoloop is a simple, convenient and effective method to cryopreserve the sperm, and cryoloop is a hopeful freezing carrier for testicular or epididymal sperm.