Strussmann CA, Nakatsugawa H, Takashima F, Hasobe M, Suzuki T, Takai R
Department of Aquatic Biosciences, Department of Food Science and Technology, Tokyo University of Fisheries, Konan 4-5-7, Minato, Tokyo, 108-8477, Japan.
Cryobiology. 1999 Nov;39(3):252-61. doi: 10.1006/cryo.1999.2208.
The toxicity of the cryoprotectant dimethyl sulfoxide (Me(2)SO) to isolated blastomeres was examined in three fish species representative of distinct environments: marine (whiting, Sillago japonica); estuarine (pejerrey, Odontesthes bonariensis); and freshwater (medaka, Oryzias latipes). The effects of embryonic stage, Me(2)SO concentration, and cooling rate on the cryopreservation of blastomeres were also studied. Whiting sheds small planktonic eggs whereas the other two species shed large demersal eggs. Isolated blastomeres from the three species tolerated Me(2)SO concentrations up to 9% relatively well for over 5 h but lost viability rapidly at 18%. Cells from later embryonic stages (512 or 1024 cells) were more tolerant of Me(2)SO than those from earlier stages (128 or 256 cells). The three factors examined, alone or in combination, had a significant effect on the survival of blastomeres after freezing and thawing, but the extent of the effect and the optimum conditions varied with the species. In general, the highest rates of successful cryopreservation were observed with older rather than younger blastomeres, slower rather than faster cooling, and with 9-18% rather than 0% Me(2)SO. Survival rates for blastomeres cryopreserved under the most effective combination of the three factors examined for each species were 19.9 +/- 10.1% for whiting, 34.1 +/- 8.5% for medaka, and 67.4 +/- 12.8% for pejerrey. Copyright 1999 Academic Press.
在代表不同环境的三种鱼类中检测了冷冻保护剂二甲基亚砜（Me(2)SO）对分离的卵裂球的毒性：海洋鱼类（银鲈，日本银鲈）；河口鱼类（佩氏南美无须鳕，博氏南美无须鳕）；以及淡水鱼类（青鳉，青鳉属）。还研究了胚胎阶段、Me(2)SO浓度和降温速率对卵裂球冷冻保存的影响。银鲈产小型浮游卵，而另外两种鱼产大型沉性卵。来自这三种鱼类的分离卵裂球在高达9%的Me(2)SO浓度下能相对较好地耐受5小时以上，但在18%时活力迅速丧失。来自胚胎后期阶段（512或1024个细胞）的细胞比早期阶段（128或256个细胞）的细胞对Me(2)SO更具耐受性。所研究的这三个因素单独或组合使用，对冻融后卵裂球的存活有显著影响，但影响程度和最佳条件因物种而异。一般来说，观察到成功冷冻保存率最高的是较老而非较年轻的卵裂球、较慢而非较快的降温速率，以及9 - 18%而非0%的Me(2)SO。在针对每个物种所研究的三个因素的最有效组合下冷冻保存的卵裂球的存活率，银鲈为19.9 +/- 10.1%，青鳉为34.1 +/- 8.5%，佩氏南美无须鳕为67.4 +/- 12.8%。版权所有1999年学术出版社。
