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Updated: Jul 10, 2026
Isolation of Lamina Propria Mononuclear Cells from Murine Colon Using Collagenase E
Published on: September 26, 2019
Isolation and subsequent analysis of murine lamina propria mononuclear cells from colonic tissue
Benno Weigmann1, Ingrid Tubbe, Daniel Seidel
1I. Med. Clinic, Immunological Laboratory, Johannes-Gutenberg University, Obere Zahlbacherstrasse 63, 55131 Mainz, Germany. bweigm@uni-mainz.de
Insights
This study details a protocol for isolating and characterizing colonic lamina propria (LP) cells from mice. This method provides high-quality cells for investigating gut immunity and inflammatory bowel diseases.
Area of Science:
- Immunology
- Gastroenterology
- Cell Biology
Background:
- Colonic lamina propria (LP) cells are crucial for understanding gut immune responses and inflammatory bowel diseases (IBD), including Crohn's disease and ulcerative colitis.
- Accurate isolation of viable LP cells is essential for reliable downstream analyses.
Purpose of the Study:
- To present a detailed protocol for the isolation and quality control of murine colonic LP cells.
- To ensure the suitability of isolated cells for subsequent molecular and cellular investigations.
Main Methods:
- Murine colonic tissue digestion using enzymatic treatments (collagenase, DNase, dispase).
- Purification of lamina propria cells via Percoll gradient centrifugation.
- Quality control assessment including Trypan Blue exclusion for cell viability and flow cytometry for apoptosis analysis.
Main Results:
- Successful isolation of viable colonic LP cells within a 6-7 hour timeframe.
- Characterization of isolated cells confirms their suitability for further research.
Conclusions:
- This protocol provides a reliable method for obtaining high-quality murine colonic LP cells.
- The isolated cells are appropriate for various downstream applications such as gene expression, proliferation, and protein analyses in the context of gut immunity and IBD research.
Abstract:
Studies on colonic cells in the lamina propria (LP) of mice are important for understanding the cellular and immune responses in the gut, especially in inflammatory bowel diseases (such as morbus crohn and colitis ulcerosa). This protocol details a method to isolate LP cells and characterize freshly isolated cells by quality control experiments to obtain cells that can be used for further investigations. After different steps of digestion of the tissue using collagenase, DNase and dispase, the resulting cells are purified using Percoll gradient. The success of the isolation can be analyzed by cell viability test (Trypan Blue exclusion test) and by flow cytometric analysis to assess apoptosis. Finally, the isolated cells can be used for further investigations like comparative studies of mRNA expression, cell-proliferation assay or protein analysis. This protocol can be completed within 6-7 h.
