中国水产科学2024，Vol.31Issue(7)：745-753,9.DOI:10.12264/JFSC2024-0111
解冻温度和冷冻保护剂对中华鲟精原干细胞超低温冷冻保存效果的影响
Effects of thawing temperature and cryoprotectant on the ultra-low temperature cryopreservation of spermatogonial stem cells of Chinese sturgeon(Acipenser sinensis)
摘要
Abstract
The Chinese sturgeon(Acipenser sinensis)is one of the key aquatic animals under first-degree protection in China,but its germplasm resources,such as spermatogonial stem cells,are not well protected by cryoprotection technology.In this study,we investigated the effects of thawing temperature,antioxidants,and antifreeze proteins on the cryopreservation of spermatogonial stem cells of Chinese sturgeon based on the formulation of the spermatogonial stem cell cryopreservation solution used for the American paddlefish(Polyodon spathula).The aim was to establish a highly efficient method for ultra-low-temperature cryopreservation of spermatogonial stem cells of Chinese sturgeon.The best cryopreservation effect was achieved at a thawing temperature of 25℃ among the temperatures tested(20,25,30,and 40℃).The number of cells after thawing was(3.86±0.51)×105,and the cell viability reached(96.36±0.53)％.The effects of different concentrations(500 mg/L,1000 mg/L,and 1500 mg/L)of glutathione,ascorbic acid,and α-tocopherol on the cryopreservation of spermatogonial stem cells of Chinese sturgeon were examined.The results showed that the experimental group with the addition of 1000 mg/L α-tocopherol to the cryopreservation solution had the highest number of cells(7.64±0.34)×105 after thawing,which was significantly higher than that of the other groups with antioxidants added,and the cell viability reached(92.82±0.72)％.The effects of different concentrations(0.1,1.0,and 10 μg/mL)of two types of antifreeze proteins(AFPⅠ and AFPⅢ)on the cryopreservation of spermatogonial stem cells of Chinese sturgeon were examined.The experimental group with 1.0 μg/mL AFPI had the best defrosting effect,and the number of cells obtained after defrosting was(6.85±0.19)×105,which was significantly higher than the number in the other experimental groups,and the cell viability was(86.89±0.73)％.In summary,in the present study,we obtained the optimal thawing temperature for the cryopreservation of spermatogonial stem cells of Chinese sturgeon,evaluated the effects of three types of antioxidants and antifreeze proteins on the effect of cryopreservation,and established a highly efficient cryopreservation technology for spermatogonial stem cells of Chinese sturgeon.These data provide technical support for the cryopreservation and restoration of the Chinese sturgeon's germplasm resources.关键词
中华鲟/精原干细胞/超低温冷冻保存/解冻温度/抗氧化剂/抗冻蛋白Key words
Acipenser sinensis/spermatogonial stem cell/ultra-low temperature cryopreservation/thawing temperature/antioxidant/antifreeze protein分类
农业科技引用本文复制引用
舒雪密,叶欢,仲嘉,张鸣,罗江,刘源,杜浩..解冻温度和冷冻保护剂对中华鲟精原干细胞超低温冷冻保存效果的影响[J].中国水产科学,2024,31(7):745-753,9.基金项目
国家重点研发计划项目(2021YFD120030107) （2021YFD120030107）
中央级公益性科研院所基本科研业务费专项资金资助项目(2023TD08). （2023TD08）
