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Mouse Intestinal Organoid Culture Protocol
Özüm Begüm Böke1, Ezgi Bulut-Okumuş1, Hazar Eren Soydan1
1Faculty of Engineering, Genetics and Bioengineering Department, Yeditepe University, İstanbul, Turkey.
Methods in Molecular Biology (Clifton, N.J.)
|January 8, 2025
Summary
This study presents a detailed protocol for culturing, passaging, and cryopreserving intestinal organoids derived from BALB/c mice. These methods are crucial for advancing stem cell research and organoid technology.
Area of Science:
- Stem Cell Biology
- Gastroenterology
- Regenerative Medicine
Background:
- Leucine-rich repeat-containing G-protein-coupled receptor 5 (Lgr5) is a key marker for intestinal adult stem cells.
- Lgr5+ stem cells are fundamental to modern organoid technology.
- Established protocols exist for various stem cell sources, but a specific BALB/c mouse model protocol is detailed here.
Purpose of the Study:
- To provide a comprehensive protocol for BALB/c mouse-derived intestinal organoid culture.
- To describe standardized procedures for organoid passaging.
- To outline methods for cryopreservation of intestinal organoids.
Main Methods:
- Isolation of intestinal stem cells from BALB/c mice.
- Culture of organoids using specific growth media and conditions.
- Serial passaging of established organoid cultures.
- Cryopreservation techniques for long-term storage.
Main Results:
- Successful establishment and maintenance of BALB/c mouse intestinal organoids.
- Demonstration of reliable passaging for continuous culture.
- Validation of cryopreservation for preserving organoid viability and function.
Conclusions:
- The presented protocol offers a reproducible method for generating and maintaining mouse intestinal organoids.
- This standardized approach facilitates further research in intestinal stem cell biology and disease modeling.
- The cryopreservation method ensures the long-term availability of organoid resources.
