Kaushik K, Gupta Psp, Johnson P, Krishna K, Nandi S, Mondal S, Kumar Tej J N, Bence S, Cseh S
ICAR-National Institute of Animal Nutrition and Physiology, Bengaluru, India.
Department and Clinic of Obstetrics and Reproduction, University of Veterinary Science, Budapest, Hungary.
Cryo Letters. 2022 Jan-Feb;43(1):10-17.
Vitrification increases the production of reactive oxygen species (ROS) and the antioxidants in the vitrification solution may be beneficial by reducing excessive ROS production.
To evaluate the effect of retinol supplementation in vitrification solution on viability, apoptosis and development-related gene expression in vitrified sheep preantral follicles.
Preantral follicles were isolated and randomly assigned into one of five groups: Group1, control fresh preantral follicles; Group 2, vitrification treatment; Group 3, vitrification + 2 μM retinol; Group 4, vitrification + 5 μM retinol; Group 5, vitrification + 10 μM retinol. Preantral follicles were placed in vitrification solutions and then plunged into liquid nitrogen (-196°C). After a week, the follicles were thawed and analyzed for follicular viability by trypan blue exclusion method and for gene expression.
Vitrification with 5 μM retinol positively affected viability in comparison with vitrification without retinol (P < 0.05). There was no significant difference in viability among the Group 1, Group 2, Group 3 and Group 5. Expression of apoptotic genes BAX and Casp 3 were higher in the vitrified group, and vitrification with 5 μM retinol (Group 4) is comparable to the control fresh. Expressions of other apoptosis-related genes (i.e., BCL2L1, BAD and BAK) showed significant difference between the control fresh group and the vitrification group with 5 μM retinol. Expression of Annexin5 was also significantly different among various groups. The expression of development competence genes GDF-9 and BMP-15 were higher (P < 0.05) in the Group vitrified with 5 μM retinol.
The supplementation of 5 μM retinol in vitrification solution was beneficial for the vitrification of ovine preantral follicles.
玻璃化冷冻会增加活性氧（ROS）的产生，而玻璃化溶液中的抗氧化剂可能通过减少过量的ROS产生而有益。
评估在玻璃化溶液中添加视黄醇对玻璃化处理的绵羊腔前卵泡活力、凋亡及发育相关基因表达的影响。
分离腔前卵泡并随机分为五组：第1组，新鲜腔前卵泡对照；第2组，玻璃化处理；第3组，玻璃化 + 2 μM视黄醇；第4组，玻璃化 + 5 μM视黄醇；第5组，玻璃化 + 10 μM视黄醇。将腔前卵泡置于玻璃化溶液中，然后投入液氮（-196°C）。一周后，解冻卵泡，通过台盼蓝排斥法分析卵泡活力并检测基因表达。
与无视黄醇的玻璃化处理相比，添加5 μM视黄醇的玻璃化处理对卵泡活力有积极影响（P < 0.05）。第1组、第2组、第3组和第5组之间的活力无显著差异。凋亡基因BAX和Casp 3在玻璃化组中的表达较高，添加5 μM视黄醇的玻璃化处理（第4组）与新鲜对照相当。其他凋亡相关基因（即BCL2L1、BAD和BAK）的表达在新鲜对照组和添加5 μM视黄醇的玻璃化组之间存在显著差异。Annexin5的表达在各组之间也有显著差异。添加5 μM视黄醇的玻璃化组中发育能力基因GDF - 9和BMP - 15的表达较高（P < 0.05）。
在玻璃化溶液中添加5 μM视黄醇有利于绵羊腔前卵泡的玻璃化冷冻。
