Department of Anatomy and Embryology, Leiden University Medical Center; The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center;
Department of Anatomy and Embryology, Leiden University Medical Center.
J Vis Exp. 2024 Aug 16(210). doi: 10.3791/66797.
The ovarian surface epithelium (OSE), the outermost layer of the ovary, undergoes rupture during each ovulation and plays a crucial role in ovarian wound healing while restoring ovarian integrity. Additionally, the OSE may serve as the source of epithelial ovarian cancers. Although the OSE regenerative properties have been well studied in mice, understanding the precise mechanism of tissue repair in the human ovary remains hampered by limited access to human ovaries and suitable in vitro culture protocols. Tissue-specific organoids, miniaturized in vitro models replicating both structural and functional aspects of the original organ, offer new opportunities for studying organ physiology, disease modeling, and drug testing. Here, we describe a method to isolate primary human OSE (hOSE) from whole ovaries and establish hOSE organoids. We include a morphological and cellular characterization showing heterogeneity between donors. Additionally, we demonstrate the capacity of this culture method to evaluate hormonal effects on OSE-organoid growth over a 2-week period. This method may enable the discovery of factors contributing to OSE regeneration and facilitate patient-specific drug screenings for malignant OSE.
卵巢表面上皮（OSE）是卵巢的最外层，在每次排卵时都会破裂，在恢复卵巢完整性的同时对卵巢伤口愈合起着至关重要的作用。此外，OSE 可能是上皮性卵巢癌的来源。尽管 OSE 的再生特性在小鼠中已经得到了很好的研究，但由于获取人类卵巢的途径有限和合适的体外培养方案，人类卵巢组织修复的确切机制仍难以理解。组织特异性类器官是一种微型化的体外模型，可复制原始器官的结构和功能方面，为研究器官生理学、疾病建模和药物测试提供了新的机会。在这里，我们描述了一种从整个卵巢中分离原代人 OSE（hOSE）并建立 hOSE 类器官的方法。我们包括了一种形态和细胞特征分析，显示了供体之间的异质性。此外，我们还证明了这种培养方法能够在 2 周的时间内评估激素对 OSE-类器官生长的影响。这种方法可能有助于发现促进 OSE 再生的因素，并为恶性 OSE 患者进行特定药物筛选提供便利。
