Isolation and Subsequent Analysis of Murine Lamina Propria Mononuclear Cells from Colonic Tissue

Weigmann B, Tubbe I, Seidel D, Nicolaev A, Becker C, Neurath MF: Isolation and subsequent analysis of murine lamina propria mononuclear cells from colonic tissue. Nature protocols 2007, 2(10):2307-2311.

Studies on colonic cells in the lamina propria (LP) of mine 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 isolate 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 students of mRNA expression, cell-proliferation assay or protein analysis. This protocol can be completed within 6-7 h.

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