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Co-Culture of Murine Small Intestine Epithelial Organoids with Innate Lymphoid Cells
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Optimized Protocol for Characterization of Mouse Gut Innate Lymphoid Cells
Ana Valle-Noguera1, María José Gómez-Sánchez1,2, Mathilde J H Girard-Madoux3
1Department of Immunology, School of Medicine, Complutense University of Madrid, 12 de Octubre Health Research Institute (imas12), Madrid, Spain.
Frontiers in Immunology
|December 17, 2020
Summary
This study presents an optimized protocol for isolating and analyzing innate lymphoid cells (ILCs) from mouse gut tissue. The method ensures high yield and viability for accurate flow cytometry identification of ILC subsets and their cytokine production.
Area of Science:
- Immunology
- Cell Biology
Background:
- Innate lymphoid cells (ILCs) are crucial for innate immunity, found in mucosal tissues and classified into five types.
- ILCs regulate homeostasis, pathogen defense, inflammation, and tissue remodeling.
- Current ILC isolation protocols are often lengthy and yield insufficient cells for analysis.
Purpose of the Study:
- To establish a detailed protocol for obtaining single-cell suspensions highly enriched in viable ILCs from mouse gut.
- To enable accurate identification and quantification of ILC subsets using flow cytometry.
- To facilitate the study of cytokine production by ILC3s involved in gut barrier integrity.
Main Methods:
- Development of a protocol for single-cell suspension enrichment from mouse small intestine and colon.
- Application of a specific cell marker panel and flow cytometry gating strategy.
- Inclusion of a procedure for analyzing cytokine production by ILC3s.
Main Results:
- An efficient method for obtaining enriched, viable ILC samples from mouse gut lamina propria.
- Detailed cell marker panel and gating strategies for comprehensive ILC subset analysis.
- A protocol enabling the study of ILC3-mediated cytokine responses.
Conclusions:
- The presented protocol offers an efficient approach for studying mouse ILCs in the gut.
- This method provides a valuable tool for future research on ILCs and gut immunity.
- Optimized ILC isolation and analysis are critical for understanding their roles in health and disease.
