Guedes Janaína de Souza, Rodrigues Jhenifer Kliemchen, Campos Ana Luisa Menezes, Moraes Camila Cruz de, Caetano João Pedro Junqueira, Marinho Ricardo Mello
Clínica Pró-Criar - Reproductive Medicine, Belo Horizonte, MG, Brasil.
Post-graduation, Faculdade de Ciências Médicas de Minas Gerais, Belo Horizonte, MG, Brasil.
Rev Bras Ginecol Obstet. 2017 Nov;39(11):614-621. doi: 10.1055/s-0037-1606129. Epub 2017 Aug 31.
The present study aimed to evaluate the impact of vitrification on the viability of follicles using a three-dimensional (3D) in vitro culture. Bovine ovarian tissue samples ( = 5) obtained from slaughterhouses were utilized. The cortex was cut into small fragments of 2 × 3 × 0.5 mm using a tissue slicer. From these fragments, secondary follicles were first isolated by mechanical and enzymatic methods, then encapsulated in alginate gel and individually cultured for 20 days. Additional fragments of the same ovarian tissue were vitrified in a solution containing 25% glycerol and 25% ethylene glycol. After warming, the follicles underwent the same follicular isolation process that was performed for the fresh follicles. A total of 61 follicles were isolated, 51 from fresh ovarian tissue, and 10 from vitrified tissue. After the culture, the vitrified and fresh follicles showed 20% and 43.1% survival rates respectively ( = 0.290), with no significant differences. At the end of the culture, there were no significant differences in follicular diameter between the vitrified (422.93 ± 85.05 µm) and fresh (412.99 ± 102.55 µm) groups ( = 0.725). Fresh follicles showed higher mean rate of antrum formation when compared with vitrified follicles (47.1% and 20.0% respectively), but without significant difference ( = 0.167). The follicles were able to develop, grow and form antrum in the 3D system after vitrification, despite the lower results obtained with the fresh tissue.
本研究旨在利用三维（3D）体外培养评估玻璃化对卵泡活力的影响。使用从屠宰场获取的牛卵巢组织样本（n = 5）。用组织切片机将皮质切成2×3×0.5毫米的小碎片。首先通过机械和酶法从这些碎片中分离出次级卵泡，然后将其包封在藻酸盐凝胶中并单独培养20天。将同一卵巢组织的其他碎片在含有25%甘油和25%乙二醇的溶液中进行玻璃化处理。复温后，对卵泡进行与新鲜卵泡相同的卵泡分离过程。总共分离出61个卵泡，其中51个来自新鲜卵巢组织，10个来自玻璃化组织。培养后，玻璃化卵泡和新鲜卵泡的存活率分别为20%和43.1%（P = 0.290），无显著差异。培养结束时，玻璃化组（422.93±85.05微米）和新鲜组（412.99±102.55微米）的卵泡直径无显著差异（P = 0.725）。与玻璃化卵泡相比，新鲜卵泡的平均卵泡腔形成率更高（分别为47.1%和20.0%），但无显著差异（P = 0.167）。尽管新鲜组织的结果更好，但玻璃化后的卵泡在3D系统中仍能够发育、生长并形成卵泡腔。
