刀鲚精子超低温冷冻保存技术的研究

Study on sperm cryopreservation of Japanese grenadier anchovy Coilia ectenes

本文对刀鲚精子的超低温冷冻保存技术进行了研究。以解冻后精子的运动力为参数,分别探讨了不同的稀释液、不同种类不同浓度的保护剂、不同的平衡时间以及不同的稀释比例对刀鲚精子进行超低温冷冻保存的保护效果。结果显示:采用D-15稀释液,将精液和稀释液按1∶2稀释,4℃平衡20 min,加入10%DMSO,混匀后液氮面上方6 cm处平衡10 min,接着在液氮面上平衡5 min,最后投入液氮中保存,一周后,液氮蒸气中平衡5 min,37℃水浴解冻,活力效果最佳,达70%左右。

The technique of cryopreservation of sperm in Japanese grenadier anchovy Coilia ectenes was studied. The success of sperm cryopreservation was evaluated by post-thaw sperm motility. To explore methods for sperm cryopreservation, this study evaluated the effects of different extenders, cryoprotectants, their volume fractions, equilibration time and dilution ratios. The experiment results demonstrated the optimal freezing procedures for Japanese grenadier anchovy sperm is diluting the semen in D-15 extender at the ratio of 1：2, chilling it to 4℃ for 20 min, adding ME2 SO to a final concentration of 10%（v/v）, then transferring the semen in cryotubes,holding the cryotubes for 10 min at 6 cm（about-180℃） above the surface of liquid nitrogen, for 5 min on the surface of liquid nitrogen, and finally plunged into liquid nitrogen for a week. After thawed at 37℃, the sperm had the highest post-thaw motility（70%）.