Renji Hospital, Key Laboratory for the Genetics of Developmental & Neuropsychiatric Disorders (Ministry of Education), Bio-X Institutes, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200240, China.
Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, 200241, China.
Biomaterials. 2021 Dec;279:121213. doi: 10.1016/j.biomaterials.2021.121213. Epub 2021 Oct 21.
In vitro production of oocytes capable of producing offspring has exciting potential applications in reproductive medicine. Here, we generated and characterized an ovarian organoid model derived from female germline stem cells using a three-dimensional culture system. We show that this model generated normal offspring and detected drug toxicity. The ovarian organoids could produce oocytes and exhibited endocrine functions. Single-cell analysis of ovarian organoids identified six ovarian cell lineages, such as germ, granulosa and theca cells, and produced gene-expression signatures for each cell type. Investigation of the expression patterns of genes related to meiosis and gene ontogeny analysis for germ cell clusters showed that a germ cell population was maintained in the ovarian organoids. Moreover, flow cytometric analysis confirmed that the population of germ cells could be maintained on the organoids and showed that ascorbic acid treatment had a beneficial effect of germ cell population maintenance on the organoids. Furthermore, we demonstrated the successful production of offspring from oocytes derived from ovarian organoids. Finally, we showed the ovarian organoids had the potential to drug toxicological detection. For example, we found that salinomycin impaired the formation of ovarian organoids and germ cell population maintenance by inducing apoptosis. These results indicate that the female germline stem cell-derived ovarian organoids represent a valuable model system for generating oocytes that can yield offspring, and provide a novel model for drug screening and toxicological detection.
体外产生能够产生后代的卵子在生殖医学中有令人兴奋的潜在应用。在这里，我们使用三维培养系统从雌性生殖干细胞中生成和表征了一种卵巢类器官模型。我们表明，该模型产生了正常的后代并检测到了药物毒性。卵巢类器官可以产生卵子并表现出内分泌功能。对卵巢类器官的单细胞分析鉴定了六个卵巢细胞谱系，如生殖细胞、颗粒细胞和膜细胞，并为每种细胞类型生成了基因表达特征。对与减数分裂和基因本体分析相关的基因表达模式的研究表明，卵巢类器官中维持着一个生殖细胞群体。此外，流式细胞术分析证实，生殖细胞群体可以在类器官上维持，并表明抗坏血酸处理对类器官上生殖细胞群体的维持有有益的影响。此外，我们证明了可以从卵巢类器官中产生的卵子产生后代。最后，我们表明卵巢类器官具有进行药物毒理学检测的潜力。例如，我们发现，博来霉素通过诱导细胞凋亡，破坏卵巢类器官的形成和生殖细胞群体的维持。这些结果表明，雌性生殖干细胞衍生的卵巢类器官代表了一种有价值的模型系统，可用于产生能够产生后代的卵子，并为药物筛选和毒理学检测提供了一种新的模型。
