Abstract
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.
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Acknowledgments
We thank Lisa Speck, Caterina Poeppel, Petra Reckling, and Bianka Drawert, Leibniz Institute for Farm Animal Biology (FBN) Dummerstorf, Germany, for excellent technical assistance. We appreciate Dagmar Viertel, Leibniz Institute for Zoo and Wildlife research (IZW) Berlin, Germany, for providing electron microscopy pictures. The authors are active members of the COST Action CA16119 (In vitro 3D total cell guidance and fitness). The original work mentioned in this chapter was partially supported by grants from the German Research Foundation (DFG Scho1231/7-1; CH2321/1-1). The Authors are members of the COST Action CA16119 In vitro 3-D total cell guidance and fitness (CellFit).
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Chen, S., Schoen, J. (2021). Using the Air–Liquid Interface Approach to Foster Apical–Basal Polarization of Mammalian Female Reproductive Tract Epithelia In Vitro. In: Brevini, T.A., Fazeli, A., Turksen, K. (eds) Next Generation Culture Platforms for Reliable In Vitro Models . Methods in Molecular Biology, vol 2273. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1246-0_18
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DOI: https://doi.org/10.1007/978-1-0716-1246-0_18
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