中间球海胆精子的超低温保存技术

Study on Spermatozoa Cryopreservation of Sea Urchin, Strongylocentrotus intermedius

20 0 0年 10～ 11月 ,对大连龙王塘海区成熟的中间球海胆 (Strongylocentrotusintermedius)精子进行超低温保存研究。结果表明 :不同的抗冻保护剂 ,不同的体积分数 ,不同的冷冻方式 ,精子冻存的效果不同。以自然海水为基础溶液 ,以 12 %的二甲亚砜或 9%的甘油作为抗冻保护剂 ,稀释精液至 8× 10 7～ 11× 10 7个 /ml,距液氮表面 10cm处预冷 8min ,再浸入液氮中保存 ,解冻时直接移出液氮于 17～ 18℃自来水流水解冻 ,精子存活率分别是 95 % ,90 % ;受精率分别为 94 0 % ,86 5 %。

Spermatozoa cryopreservation of mature sea urchin, Strongylocentrotus intermedius ,from the sea beach of Long wan tang in Dalian was studied in 2000 Based on the solution of natural sea water,used 12%DMSO or 9%,glycerine as the cryoprotector,the density of the diluent spermatozoa were 8×10 7～11×10 7inds/ml.The results of freezing and thawing tests showed that survival rate of the spermatozoa were 95%,90%,the fertilization rate were 94 0%,86 5% when spermatozoa were maintained at 10 cm above LN 2 for 8 min before load into LN 2,and thawed in tap water(17～ 18℃)