Emory University, Atlanta, Georgia

3 ARTICLES PUBLISHED IN JoVE

Biology

Cryosectioning Method for Microdissection of Murine Colonic Mucosa

Attila E. Farkas¹, Christian Gerner-Smidt¹, Loukia Lili¹, Asma Nusrat¹, Christopher T. Capaldo¹

¹Epithelial Pathobiology and Mucosal Inflammation Research Unit, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia

Here we describe a simple method for the isolation of spatially distinct murine colonic epithelial surface cells from the underlying crypt-base cells.

Immunology and Infection

Systematic Scoring Analysis for Intestinal Inflammation in a Murine Dextran Sodium Sulfate-Induced Colitis Model

Vicky Garcia-Hernandez¹, Philipp-Alexander Neumann², Stefan Koch^3,4, Renae Lyons¹, Asma Nusrat¹, Charles A. Parkos¹

¹Department of Pathology, University of Michigan, ²Department of Surgery, Klinikum Rechts der Isar, Technische Universität München, ³Wallenberg Centre for Molecular Medicine, Linköping University, ⁴Department of Biomedical and Clinical Sciences (BKV), Linköping University

Systematic scoring of intestinal inflammation using a free computer-assisted system is a powerful tool to quantitatively compare histopathological changes in colitis models characterized by the presence of ulcers and inflammatory changes. Histological colitis score evaluation strengthens clinical observations and facilitates data interpretation.

Biology

Generation of Murine Primary Colon Epithelial Monolayers from Intestinal Crypts

Chithra K Muraleedharan¹, Jay Mierzwiak¹, Darius Feier¹, Asma Nusrat¹, Miguel Quiros¹

¹Department of Pathology, School of Medicine, University of Michigan

In this protocol, we describe how to generate murine primary epithelial colon monolayers directly from intestinal crypts. We provide experimental approaches to generate confluent monolayers on permeable filters, confluent monolayers for scratch wound healing and biochemical studies, and sparse and confluent monolayers for immunofluorescence analysis.