摘要
以牙鲆胚胎为材料，对胚胎程序化冷冻保存的主要环节进行了探讨，以优化牙鲆胚胎的程序化超低温冷冻保存法。单一抗冻荆的毒性试验表明甲醇（Methanol）、1，2-丙二醇（PG）的毒性低于其他抗冻剂的毒性。混合抗冻剂的毒性试验表明，20％～25％PM抗冻剂的毒性很低，经其平衡处理的胚胎，冷冻至-30℃，成活率可达80％以上。植冰前采用2℃／min的降温速率，从植冰后到入液氮前温度采用8℃／min的降温速率效果较好。入液氮前温度的筛选实验表明，入液氮前的温度以-45℃较好。利用20％PM和22％PMP，采取优化的降温程序冷冻保存尾芽期的牙鲆胚胎，分别获得5枚和7枚复活胚胎，并全部孵化出膜。
Study was carried out on cryopreservation of Japanese flounder （Paralichthys olivaceus） embryos, aiming at optimizing the programmed freezing method. Through toxicity tests of cryoprotectant in Japanese flounder embryos, methanol （MeOH） and propylene glycol （PG） were found to be lower than other cryoprotectants in toxicity to embryos. Furthermore, the mixed cryoprotectants, viz. 20% -25 %PM （PG＋MeOH）, was even weaker in toxicity, compared to single cryoprotectant. In the cooling process, it was better to adopt 2℃/min as the velocity of descending temperature before icing, and then using 8 ℃/min. until exposing to liquid nitrogen, with reaching -45 ℃ being preferable. By using the optimized descending temperature process, 5 embryos survived in 26 flounder embryos at tail-bud stage undergoing cryopreservation with 20 %PM solution, and 7 survived in 45 with 22 % PMP solution.
出处
《海洋水产研究》
CSCD
北大核心
2007年第3期57-63,共7页
Marine Fisheries Research
基金
国家863高技术研究与发展项目(2001AA621100)
国家自然科学基金项目(30570259)共同资助
关键词
牙鲆
胚胎
程序化
冷冻保存
Paralichthys olivaceus Embryos A programmed freezing method Cryopreservation
