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Optimized protocol for dissociation of mouse intestinal epithelium to viable single-cells
Ni Zhao1, Peiling Geng1, Yong Ge1
1Department of Microbiology, Immunology & Molecular Genetics, University of Texas Health San Antonio, San Antonio, TX, USA.
Insights
This study presents a new protocol for dissociating mouse intestinal epithelial cells (IECs) using TrypLE Express and Dispase II. The method efficiently recovers millions of viable IECs, including rare enterochromaffin (EC) cells, for functional profiling.
Area of Science:
- Gastroenterology and Cell Biology
- Microbiome Research
- Host-Diet Interactions
Background:
- Intestinal epithelial cells (IECs) are vital for host-microbe and host-diet interactions.
- IECs consist of diverse cell types, making functional profiling challenging.
- Understanding IEC heterogeneity is key to deciphering gut health and disease.
Purpose of the Study:
- To develop and present a robust protocol for dissociating mouse intestinal epithelium.
- To enable efficient recovery of viable intestinal epithelial cells (IECs) for downstream analysis.
- To facilitate the isolation of rare IEC populations, such as enterochromaffin (EC) cells.
Main Methods:
- A novel protocol utilizing TrypLE Express and Dispase II for tissue dissociation.
- Detailed steps for mouse intestinal tissue collection, crypt isolation, and enzymatic digestion.
- Optimization of enzymatic digestion for maximal recovery of viable single IECs.
Main Results:
- Successful dissociation of mouse intestinal epithelium yielding millions of viable IECs.
- Demonstrated efficiency in isolating rare cell populations, specifically enterochromaffin (EC) cells.
- Protocol provides a scalable method for IEC isolation from the mouse intestine.
Conclusions:
- The presented protocol offers a reliable method for obtaining large numbers of viable mouse IECs.
- This technique is particularly valuable for studying rare cell types and their functions.
- Enables advanced functional profiling of intestinal epithelial heterogeneity in host-microbe-diet interactions.
Abstract:
The functional profiling of intestinal epithelial cells (IECs), which comprise multiple heterogeneous populations, is crucial for understanding host interactions with dietary and microbial factors. Here, we present a protocol for dissociating mouse epithelium based on TrypLE Express and Dispase II. We describe steps for tissue collection, crypt isolation, and enzyme digestion. This protocol enables the recovery of millions of viable IECs from the mouse intestine and is particularly useful for isolating rare cell populations, including enterochromaffin (EC) cells.
