小鼠微量圆形精子细胞冷冻保存方法研究

An optimal method for cryopreservation of microamount round spermatids of the mouse

目的:探讨小鼠微量圆形精子细胞的冷冻方法和条件。方法:比较玻璃化冷冻和常规慢冷冻,不同浓度冷冻保护剂(5%、7%、9%甘油)及不同平衡时间(0、15、30、45、60 min)对体外培养所获微量小鼠圆形精子细胞的冷冻效果。结果:玻璃化冷冻和常规慢冷冻在7%甘油中平衡时间30 min均可获得较高的细胞复苏率,且玻璃化冷冻小鼠圆形精子细胞的复苏率显著高于常规慢冷冻方法[(72.9±15.4)%vs.(58.2±17.7)%,P<0.05]。结论:采用玻璃化冷冻方法,在7%浓度的甘油保护剂中平衡30 min,可获得较满意的微量圆形精子细胞冷冻效果。

Objective： To search for an optimal protocol and freezing conditions for the cryopreservation of microamount round spermatids of the mouse. Methods： We compared the survival rates of frozen-thawed microamount round spermatids of the mouse achieved by vitrification or standard slow freezing with different concentrations of glycerol （5,7, or 9% ） and different lengths of equi- librium time （0, 15, 30, 45, or 60 rain）. Results： Under the conditions of 7% glycerol and 30 rain equilibrium, both vitrification and standard slow freezing achieved high survival rates of spermatids, and the former obtained an even higher rate than the latter （[72.9±15.41% vs [58.2 ±17.71%, P〈0.05）. Conclusion： A high rate of frozen-thawed microamount round spermatids of the mouse can be achieved by vitrification under the conditions of 7% glycerol and 30 min equilibrium.