栉孔扇贝胚胎超低温液氮保存

CRYOPRESERVATION OF EMBRYOS OF SCALLOP, Chlamys farreri, IN LIQUID NITROGEN

选用两种低温保护剂配方，Ａ；１０％甘油＋４％葡萄糖；Ｂ：１５％ＤＭＳＯ（二甲 基亚铜）＋４％葡萄糖＋２％柠檬酸盐，利用微机控制生物降温仪，对栉孔扇贝胚胎 进行液氮低温保存。以１℃／ｍｉｎ由室温降温至－２０℃，接着以２０℃／ｍｉｎ降温至－ ８０℃，然后样品直接入液氮保存（－１９６℃），１周后，经４５℃水浴解冻，栉孔扇贝胚 胎（发育 ６ｄ）的存活率达 ６５． １％（Ａ配方）和 ５６． ６％（Ｂ配方）；而采用６℃／ｍｉｎ降温 至－２０℃，并以 ２０℃／ｍｉｎ降温至－８０℃，直接入液氮，胚胎解冻后的存活率分别为 ３９．０％（Ａ配方）和２７．９％（Ｂ配方）。选用发育２４ｈ的栉孔扇贝胚胎进行液氮保存 实验，解冻后，胚胎基本死亡。

Two cryoprotective recipes, A: 10% glycerol+4% glucose, B: 15% DMSO (dimethl sulfoxide ) + 4 % glucose + 2 % citrate, were used in cropreservation of embryos of Chinese scallop,Chlamys farreri,by the aid of Programmed Cryobiological Controller. Using the freezing procedure,i. e. 1℃ /min to - 20℃, 20℃/min to - 80℃, then plunging the samples into liquid nitrogen, the scallop embryos (6d, veliger) survival rate can reach as high as 65. 1 % (recipe A) and 56. 5% (recipe B) after thawing in 45℃ sea wa- ter bath. But the freezing procedure, 6℃/the to - 20℃ - 20℃/nd to - 80℃, them plunging the samples into liquid nitrogen, presented comparatively low embryo (6d)survival rate, i, e. 39. 0% (recipe A) and 27. 9% (recipe B). No embryo survived when cryopreserving the 24h embryo (trochophore)in liquid nitrogen.