Separation of Avian Preovulatory Follicle Granulosa and Theca Cell Layers for Downstream Applications
| Author(s) | Kramer, Ashley E. | |
| Author(s) | Ellwood, Kathryn M. | |
| Author(s) | Brannick, Erin M. | |
| Author(s) | Dutta, Aditya | |
| Date Accessioned | 2025-01-14T17:44:20Z | |
| Date Available | 2025-01-14T17:44:20Z | |
| Publication Date | 2024-10-24 | |
| Description | This article was originally published in Journal of Visualized Experiments. The version of record is available at: https://doi.org/10.3791/67344. Copyright © 2024 JoVE Journal of Visualized Experiments. This article will be embargoed until 10/25/2026. | |
| Abstract | Summary Here, we describe a protocol for separating yolk, granulosa cells, and theca cells in avian preovulatory follicles. This precision handling enables critical investigations into the role of these layers in reproductive function, aiding the understanding of follicular development, hormonal regulation, and disease research for enhanced agricultural yield and biomedical insights. Abstract Layer hens (egg-laying chickens) and broiler breeders (breeding stock for meat-producing chickens) are crucial to the world's food supply as a reliable source of protein. They are also an emerging animal model for the study of human reproductive disease. As the field of poultry research develops, the health and function of the layer hen and broiler breeder ovary will be an important point of study for both agricultural and biomedical researchers. One of the challenges presented by this emerging interest is the need for replicable techniques that all researchers can employ in ovarian specimen collection. In particular, a detailed visual process must be established to define the proper separation of the specialized granulosa and theca cell layers from hen follicles to achieve agreement and consistency among researchers. This study describes the extraction of preovulatory follicles and ovary tissue in white leghorn hens of prime reproductive age. The separation of these follicles is performed under cold, liquid conditions to congeal the yolk for easier manipulation and to prevent the follicle's own weight from tearing apart cell layers during the separation process. Once the separation is complete, the desired cell layers can be further digested for tissue culture approaches or can be cryopreserved for genomic and proteomic analyses. | |
| Sponsor | We are grateful to Milos Markis (AviServe) for assistance with animal husbandry, Nicole Guarino (University of Delaware) for assistance with manuscript preparation, Evelyn Weaver and Ramesh Ramachandran (The Pennsylvania State University) for assistance with procedure demonstration and manuscript preparation. Figure 2A and Figure 3 were created using BioRender.com using an institutional license sponsored by the University of Delaware Research Office. This work was supported by the UD CANR Comparative Pathology Laboratory. KME is supported by USDA NIFA grant 2023-67011-40333. This work was supported by grants from the University of Delaware Research Foundation (UDRF) and the Delaware INBRE program (supported by a grant from the National Institute of General Medical Sciences - NIGMS P20 GM103446 from the National Institutes of Health and the State of Delaware) to AD. | |
| Citation | Kramer, A. E., Ellwood, K. M., Brannick, E. M., Dutta, A. Separation of Avian Preovulatory Follicle Granulosa and Theca Cell Layers for Downstream Applications. J. Vis. Exp. (212), e67344, doi:10.3791/67344 (2024). | |
| ISSN | 1940-087X | |
| URL | https://udspace.udel.edu/handle/19716/35718 | |
| Language | en_US | |
| Publisher | Journal of Visualized Experiments | |
| Title | Separation of Avian Preovulatory Follicle Granulosa and Theca Cell Layers for Downstream Applications | |
| Type | Article |
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