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Updated: Jul 17, 2026
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Generation of Porcine Testicular Organoids with Testis Specific Architecture using Microwell Culture
Published on: October 3, 2019
Testicular Organoid Formation in Microwell Culture
Nathalia de Lima E Martins Lara1, Anja Elsenhans1, Ina Dobrinski2
1Department of Biochemistry and Molecular Biology, Cumming School of Medicine and Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, Canada.
Methods in Molecular Biology (Clifton, N.J.)
|March 27, 2025
Summary
Testicular organoids offer a 3D in vitro model for male reproduction research, aiding fertility preservation and toxicity screening. This study details a simple microwell protocol for generating these organoids from porcine testes.
Area of Science:
- Male reproductive biology
- 3D in vitro modeling
Background:
- 2D cell cultures and animal models have limitations in studying male reproduction.
- Testicular organoids provide a more physiologically relevant 3D in vitro platform.
Purpose of the Study:
- To present a simple, reproducible protocol for generating testicular organoids.
- To enable studies on testicular cell interactions, development, and the stem cell niche.
- To facilitate applications in fertility preservation, disease modeling, and toxicity screening.
Main Methods:
- Enzymatic digestion of immature testes to obtain a single-cell suspension.
- Centrifugation of cells into pyramid-shaped microwells for aggregation.
- Culture of aggregated cells to form homogenous testicular organoids.
Main Results:
- Generation of thousands of homogenous testicular organoids.
- Recapitulation of testicular cytoarchitecture and functions in vitro.
- A reproducible protocol applicable to various mammalian species.
Conclusions:
- The developed microwell-based protocol simplifies testicular organoid generation.
- Testicular organoids are valuable tools for advancing male reproductive research.
- This method supports applications in fertility, disease modeling, and reproductive toxicology.
