Nuffield Department of Women's and Reproductive Health, University of Oxford, Oxford, UK.
Department of Paediatric Oncology and Haematology, Oxford University Hospitals NHS Foundation Trust, Oxford, UK.
Reprod Fertil. 2021 Mar 23;2(1):59-68. doi: 10.1530/RAF-20-0058. eCollection 2021 Jan.
follicle growth is a potential fertility preservation method for patients for whom current methods are contraindicated. Currently, this method has only been successful using fresh ovarian tissue. Since many patients who may benefit from this treatment currently have cryopreserved ovarian tissue in storage, optimising follicle growth (IVG) for cryopreserved-thawed tissue is critical. This study sought to improve the first step of IVG by comparing different short-term culture systems for cryopreserved-thawed human ovarian tissue, in order to yield a higher number of healthy multilayer follicles. We compared two commonly used culture media (αMEM and McCoy's 5A), and three plate conditions (300 µL, 1 mL on a polycarbonate membrane and 1 mL in a gas-permeable plate) on the health and development of follicles after 6 days of culture. A total of 5797 follicles from three post-pubertal patients (aged 21.3 ± 2.3 years) were analysed across six different culture conditions and non-cultured control. All culture systems supported follicle development and there was no difference in developmental progression between the different conditions tested. Differences in follicle morphology were evident with follicles cultured in low volume conditions having significantly greater odds of being graded as morphologically normal compared to other conditions. Furthermore, culture in a low volume of αMEM resulted in the highest proportion of morphologically normal primary and multilayer follicles (23.8% compared to 6.3-19.9% depending on condition). We, therefore, recommend culturing cryopreserved human ovarian tissue in a low volume of αMEM to support follicle health and development.
Ovaries contain a large number of follicles, each containing an immature egg and other important cells. Cancer treatments can lead to long-lasting negative side effects to the ovaries including the destruction of follicles, resulting in infertility. One strategy to preserve fertility is freezing of ovaries or ovarian tissue in girls and women undergoing cancer treatment. The long-term aim is to thaw and grow their ovarian tissue in the laboratory to obtain mature eggs, which can then be fertilised. In this study, we compared six different methods of growing previously frozen human ovarian tissue in order to best support follicle growth and health. We found that using the lowest amount of αMEM medium (a specific type of nutrient-rich growth solution) resulted in the highest proportion of healthy follicles. Improving the methods used to grow ovarian tissue, particularly frozen tissue, is important for future fertility preservation.
卵泡生长是一种有生育能力的保留方法，适用于目前方法禁忌的患者。目前，这种方法仅使用新鲜卵巢组织成功。由于许多可能受益于这种治疗的患者目前在储存中冷冻保存了卵巢组织，因此优化冷冻保存-解冻组织的卵泡生长（IVG）至关重要。本研究通过比较不同的短期培养系统来改善 IVG 的第一步，以获得更多健康的多层卵泡。我们比较了两种常用的培养基（αMEM 和 McCoy's 5A），以及三种平板条件（300μL、1mL 在聚碳酸酯膜上和 1mL 在透气板上），在 6 天的培养后评估卵泡的健康和发育情况。总共对来自三个青春期后的患者（年龄 21.3±2.3 岁）的 5797 个卵泡进行了分析，这些卵泡分布在六种不同的培养条件和未培养的对照组中。所有培养系统均支持卵泡发育，并且在不同的测试条件下，发育进展没有差异。卵泡形态的差异明显，低体积条件下培养的卵泡有更高的几率被评为形态正常，与其他条件相比。此外，在低体积的αMEM 中培养导致形态正常的初级和多层卵泡的比例最高（23.8%，而取决于条件的其他比例为 6.3%-19.9%）。因此，我们建议在低体积的αMEM 中培养冷冻保存的人类卵巢组织，以支持卵泡的健康和发育。
卵巢中含有大量的卵泡，每个卵泡中都含有一个未成熟的卵子和其他重要的细胞。癌症治疗会对卵巢造成长期的负面影响，包括卵泡的破坏，从而导致不孕。一种保留生育能力的策略是在接受癌症治疗的女孩和女性中冷冻卵巢或卵巢组织。长期目标是在实验室中解冻和生长她们的卵巢组织以获得成熟的卵子，然后可以对其进行受精。在这项研究中，我们比较了六种不同的方法来生长以前冷冻的人类卵巢组织，以最好地支持卵泡生长和健康。我们发现，使用最低量的αMEM 培养基（一种特定类型的富含营养的生长溶液）会导致最高比例的健康卵泡。改善用于生长卵巢组织的方法，特别是冷冻组织的方法，对未来的生育能力保留很重要。
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