Document Detail

Isolation and subsequent analysis of murine lamina propria mononuclear cells from colonic tissue.
MedLine Citation:
PMID:  17947970     Owner:  NLM     Status:  MEDLINE    
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.
Benno Weigmann; Ingrid Tubbe; Daniel Seidel; Alex Nicolaev; Christoph Becker; Markus F Neurath
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Nature protocols     Volume:  2     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2007  
Date Detail:
Created Date:  2007-10-19     Completed Date:  2008-01-22     Revised Date:  2008-03-24    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  2307-11     Citation Subset:  IM    
I. Med. Clinic, Immunological Laboratory, Johannes-Gutenberg University, Obere Zahlbacherstrasse 63, 55131 Mainz, Germany.
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MeSH Terms
Cell Culture Techniques
Cell Separation / methods*
Colon / cytology,  immunology*
Intestinal Mucosa / cytology,  immunology*
Lymphocytes / cytology*
Tissue Culture Techniques

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