Protocol to establish turkey oviductal organoids as an in vitro model

• Published in: STAR protocols 2024-12, Vol.5 (4), p.103384, Article 103384
• Main Authors: Santativongchai, Pitchaya, Klaeui, Caitlin C., Kosonsiriluk, Sunantha, Saqui-Salces, Milena, Reed, Kent M., Wileman, Ben W., Studniski, Marissa M., Boukherroub, Kahina S.
• Format: Article
• Language: English
• Subjects: cell biology; organoids; tissue engineering
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2024.103384

The study of reproductive function in turkey hens has been difficult due to the lack of a reliable, representative in vitro model for investigating profound physiological aspects. This article presents a protocol to establish turkey oviductal organoids, including steps for isolating turkey oviduct epithelial cells followed by seeding and maintaining 3D organoid cultures. We also detail procedures for organoid fixation for histological analysis. This organoid model could serve as a valuable in vitro tool for understanding the intricacies of turkey reproductive physiology. [Display omitted] •Steps for the isolation and organoid development of turkey oviduct epithelial cells•Procedures to expand, cryopreserve, thaw, and fix the turkey oviductal organoids•Organoids preserve the expressions of progesterone and avian influenza virus receptors•Turkey infundibulum and magnum organoids could be used for long-term studies Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. The study of reproductive function in turkey hens has been difficult due to the lack of a reliable, representative in vitro model for investigating profound physiological aspects. This article presents a protocol to establish turkey oviductal organoids, including steps for isolating turkey oviduct epithelial cells followed by seeding and maintaining 3D organoid cultures. We also detail procedures for organoid fixation for histological analysis. This organoid model could serve as a valuable in vitro tool for understanding the intricacies of turkey reproductive physiology.