Meizhou Clinical Institute of Shantou University Medical College.
Department of Pathology, Meizhou People's Hospital.
J Vis Exp. 2024 Aug 2(210). doi: 10.3791/66928.
Endometrial organoids offer valuable insights into the development and pathophysiology of endometrial diseases and serve as platforms for drug testing. While human and mouse endometrial organoids have been developed, research on rat endometrial organoids remains limited. Given that rats can better simulate certain endometrial pathologies, such as intrauterine adhesions, this study aimed to establish rat endometrial organoids. We present a detailed protocol for the isolation and culture of rat endometrial epithelial stem cells (reESCs) and the generation of rat endometrial organoids. Using a refined reESCs expansion medium, we successfully isolated and stably expanded reESCs, demonstrating their long-term culture potential. The reESC-generated organoids exhibited typical structural and functional characteristics of the endometrium, including hormone responsiveness. Our results showed that rat endometrial organoids could be cultured over a long term with stable proliferation, maintaining the glandular structure, cell polarity, and functional characteristics of the endometrial epithelium. This novel rat-derived endometrial organoid model provides a valuable platform for studying endometrial diseases and testing therapeutic interventions, with potential applications across various mammalian species.
子宫内膜类器官为子宫内膜疾病的发展和病理生理学提供了有价值的见解，并可作为药物测试的平台。虽然已经开发出了人类和小鼠的子宫内膜类器官，但大鼠子宫内膜类器官的研究仍然有限。鉴于大鼠可以更好地模拟某些子宫内膜病理，如宫腔粘连，本研究旨在建立大鼠子宫内膜类器官。我们提出了一种详细的大鼠子宫内膜上皮干细胞（reESC）分离和培养以及大鼠子宫内膜类器官生成的方案。使用改良的 reESC 扩增培养基，我们成功地分离并稳定地扩增了 reESC，证明了它们具有长期培养的潜力。由 reESC 生成的类器官表现出典型的子宫内膜结构和功能特征，包括激素反应性。我们的结果表明，大鼠子宫内膜类器官可以在长期培养中稳定增殖，维持腺体结构、细胞极性和子宫内膜上皮的功能特征。这种新型大鼠来源的子宫内膜类器官模型为研究子宫内膜疾病和测试治疗干预提供了有价值的平台，具有在各种哺乳动物物种中的潜在应用。
