Leal Gabriela Ramos, Graciosa Maria Alice Guimarães, Monteiro Clara Ana Santos, Pasolini Renata, Dos Reis Camargo Agostinho Jorge, Oliveira Clara Slade, de Paula Vasconcelos Carlos Otávio, Garcia Nogueira Luiz Altamiro, Reis Ferreira Ana Maria, Serapião Raquel Varella
Universidade Federal Fluminense (UFF), Department of Veterinary Medicine, Vital Brazil Filho St., 64, 24230-340, Niteroi, Rio de Janeiro, Brazil.
Empresa de Pesquisa Agropecuária do Estado do Rio de Janeiro (PESAGRO RIO), São Boa Ventura Av., 770, 24120-19, Fonseca, Niteroi, Rio de Janeiro, Brazil.
Theriogenology. 2020 Dec;158:277-282. doi: 10.1016/j.theriogenology.2020.09.024. Epub 2020 Sep 18.
This study aimed to test the effects of an IVM SPOM adaptation (SPOM-adapted IVM) on the production, total number of cells (TNC), apoptosis, and cryotolerance (post-warming survival and cytoskeleton actin integrity) of bovine IVP embryos. Two experiments were conducted with two experimental groups based on IVM treatment: A control group (TCM 199 without FCS) and an SPOM-adapted group (TCM 199 with forskolin and IBMX in pre-IVM and IVM with cilostamide). The first experiment evaluated embryo in vitro production, TNC, and apoptosis rate on D9 of development. In the second experiment, embryos were vitrified/warmed at D7 (control fresh and vitrified; SPOM-adapted fresh and vitrified) and assessed regarding post-warming survival rates and cytoskeleton actin integrity. Statistical analysis was performed using GraphPad INSTAT software at a significance level of 5%. An increase (p < 0.05) in blastocyst production was observed in the SPOM-adapted group comparing to the control group. There was no difference (p > 0.05) in the TNC or apoptosis rate between the groups. Regarding cryopreservation, no differences were found (p > 0.05) in actin integrity or post-warming survival rates between the vitrified groups. In both vitrified groups, we observed a significantly lower uninjured pattern of actin integrity compared to the fresh groups (p < 0.05). We conclude that the SPOM-adapted IVM system is beneficial for blastocyst production and does not affect the quality and cryotolerance of the produced embryos.
本研究旨在测试体外成熟（IVM）的促卵泡素（SPOM）适应性方案（SPOM 适应性 IVM）对牛体外受精（IVP）胚胎的产量、细胞总数（TNC）、凋亡以及冷冻耐受性（解冻后存活率和细胞骨架肌动蛋白完整性）的影响。基于 IVM 处理进行了两个实验，设有两个实验组：一个对照组（不含胎牛血清的 TCM 199）和一个 SPOM 适应性组（在 IVM 前使用福司可林和异丁基甲基黄嘌呤的 TCM 199，以及在 IVM 时使用西洛他唑的 TCM 199）。第一个实验评估了发育第 9 天的胚胎体外生产情况、TNC 和凋亡率。在第二个实验中，胚胎在第 7 天进行玻璃化/解冻（对照组新鲜和玻璃化；SPOM 适应性组新鲜和玻璃化），并评估解冻后的存活率和细胞骨架肌动蛋白完整性。使用 GraphPad INSTAT 软件进行统计分析，显著性水平为 5%。与对照组相比，SPOM 适应性组的囊胚产量有所增加（p < 0.05）。两组之间的 TNC 或凋亡率没有差异（p > 0.05）。关于冷冻保存，玻璃化组之间在肌动蛋白完整性或解冻后存活率方面没有发现差异（p > 0.05）。在两个玻璃化组中，与新鲜组相比，我们观察到肌动蛋白完整性的未受损模式显著更低（p < 0.05）。我们得出结论，SPOM 适应性 IVM 系统有利于囊胚生产，并且不影响所产生胚胎的质量和冷冻耐受性。
