Institute of Reproductive Biology, Leibniz Institute for Farm Animal Biology (FBN), Dummerstorf, Germany.
Methods Mol Biol. 2021;2273:251-262. doi: 10.1007/978-1-0716-1246-0_18.
Oviduct and uterus are key female reproductive organs lined by ciliated simple columnar epithelia, which are the first line of maternal contact with gametes and the developing embryo during reproduction and which warrant the optimal developmental environment for the conceptus. A major challenge for modeling these epithelia in vitro is the preservation of apical-basal polarization and cilia formation. The air-liquid interface (ALI) culture approach is a technology originally invented for modeling epidermal and airway epithelia. It has recently been shown that it also allows the establishment of highly differentiated in vitro models of epithelia that do not have access to ambient air in vivo. In this chapter, we present a comprehensive ALI procedure to model female reproductive tract (FRT) epithelia of different mammalian species in vitro over extended time periods. As a working example, the protocol focuses on primary oviductal epithelial cells (OEC) isolated from domestic pig. Hints on protocol variations for the culture of OEC from other species are provided in the Subheading 4.
输卵管和子宫是由纤毛单层柱状上皮衬里的女性主要生殖器官，它们是母体在生殖过程中与配子和发育中的胚胎首次接触的部位，为胚胎提供了最佳的发育环境。在体外模拟这些上皮组织的一个主要挑战是保持顶底极性和纤毛形成。气液界面（ALI）培养方法是最初为模拟表皮和气道上皮而发明的一项技术。最近的研究表明，它还可以建立体内无法接触到周围空气的上皮细胞的高度分化的体外模型。在本章中，我们介绍了一种全面的 ALI 程序，用于在体外长时间内模拟不同哺乳动物物种的女性生殖道（FRT）上皮细胞。作为一个工作示例，该方案侧重于从家猪中分离的原代输卵管上皮细胞（OEC）。在标题 4 中提供了针对其他物种 OEC 培养的方案变化提示。
