Keivanloo Saeide, Sudagar Mohammad
Department of Fishery Sciences, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran.
Department of Fishery Sciences, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran.
Theriogenology. 2016 Mar 15;85(5):1013-1018. doi: 10.1016/j.theriogenology.2015.11.012. Epub 2015 Dec 2.
Vitrification could provide a promising tool for the cryopreservation of fish embryos. To achieve successful cryopreservation, several parameters should be taken into account in the design of a vitrification protocol. In the present study, some relevant factors were investigated (choice of a proper vitrificant solutions and temperature for thawing) using neurulation-stage embryos. Six DMSO-based vitrificant solutions (V1-V6) were tested using a 6-step incorporation protocol. DMSO-based vitrificant solutions contained DMSO + permeable cryoprotectants + nonpermeable cryoprotectants. Embryos were immersed in vitrificant solutions for 7 minutes and directly plunged into liquid nitrogen. After vitrification (-196 °C for 10 minutes), the thawing was performed in a water bath at 0 or 20 °C and then embryos incubated until hatched. Our results demonstrated that some embryos vitrified in 5 of 6 vitrification solutions survived and hatched out, but none survived after vitrification in V2. The highest survival rate (45.45%) was observed in samples frozen with the best vitrificant solution (V6) and thawing combination (20 °C). These results establish that cryopreservation of Persian sturgeon (Acipenser persicus) embryos by DMSO-based vitrificant solutions is possible.
玻璃化可为鱼类胚胎的冷冻保存提供一种有前景的工具。为实现成功的冷冻保存，在设计玻璃化方案时应考虑几个参数。在本研究中，使用神经胚期胚胎对一些相关因素（合适的玻璃化溶液的选择和解冻温度）进行了研究。使用六步导入方案测试了六种基于二甲基亚砜（DMSO）的玻璃化溶液（V1 - V6）。基于DMSO的玻璃化溶液包含DMSO + 渗透性冷冻保护剂 + 非渗透性冷冻保护剂。将胚胎浸入玻璃化溶液中7分钟，然后直接投入液氮。玻璃化处理（-196°C 10分钟）后，在0或20°C的水浴中进行解冻，然后将胚胎孵育至孵化。我们的结果表明，在六种玻璃化溶液中的五种中玻璃化的一些胚胎存活并孵化出来，但在V2中玻璃化后没有胚胎存活。在用最佳玻璃化溶液（V6）和解冻组合（20°C）冷冻的样本中观察到最高存活率（45.45%）。这些结果表明，使用基于DMSO的玻璃化溶液冷冻保存波斯鲟（Acipenser persicus）胚胎是可能的。
