Dextran Sulfate Sodium (DSS)‐Induced Colitis in Mice

Benoit Chassaing1, Jesse D. Aitken1, Madhu Malleshappa1, Matam Vijay‐Kumar2

1 Center for Inflammation, Immunity, & Infection, Department of Biology, Georgia State University, Atlanta, Georgia, 2 Department of Medicine, The Pennsylvania State University College of Medicine, Hershey, Pennsylvania
Publication Name:  Current Protocols in Immunology
Unit Number:  Unit 15.25
DOI:  10.1002/0471142735.im1525s104
Online Posting Date:  February, 2014
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Abstract

Inflammatory bowel diseases (IBD), mainly comprising ulcerative colitis and Crohn's Disease, are complex and multifactorial diseases with unknown etiology. For the past 20 years, to study human IBD mechanistically, a number of murine models of colitis have been developed. These models are indispensable tools to decipher underlying mechanisms of IBD pathogenesis as well as to evaluate a number of potential therapeutics. Among various chemically induced colitis models, the dextran sulfate sodium (DSS)‐induced colitis model is widely used because of its simplicity and many similarities with human ulcerative colitis. This model has both advantages and disadvantages that must be considered when employed. This protocol describes the DSS‐induced colitis model, focusing on details and factors that could affect DSS‐induced pathology. Curr. Protoc. Immunol. 104:15.25.1‐15.25.14. © 2014 by John Wiley & Sons, Inc.

Keywords: chemical colitogen; intestinal inflammation; occult blood; myeloperoxidase; ulcerative colitis; inflammatory bowel disease

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Induction and Evaluation of DSS Colitis in Mice
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Induction and Evaluation of DSS Colitis in Mice

  Materials
  • Male or female mice (8 weeks old; C57BL/6J and BALB/c)
  • 2.0% to 5.0% dextran sulfate sodium (DSS) salt (reagent‐grade, mol. wt. 36 to 50 kDa; MP Biomedicals, cat. no. 02160110) in autoclaved drinking water
  • 1.5% to 2.0% isoflurane anesthesia
  • Brominated deoxyuridine (BrdU, Invitrogen, cat. no. E10187)
  • 4 and 40 KDa FITC‐dextran (Sigma, cat. no. FD4)
  • PBS
  • 70% ethanol
  • 0.5% hexadecyltrimethylammonium bromide (Sigma, cat. no. H6269) in 50 mM PBS, pH 6.0
  • 1 mg/ml dianisidine dihydrochloride (Sigma, cat. no. D3252)
  • 0.00005% H 2O 2
  • RNAlater (Sigma, cat. no. R0901)
  • 10% buffered formalin
  • HBSS with 1.0% antibiotics (penicillin and streptomycin, Fisher, cat. no. BP2959)
  • Serum‐free RPMI1640 medium with 1.0% antibiotics (penicillin and streptomycin)
  • Mice cages
  • Ear punch
  • Animal balance
  • Sterile forceps
  • Microcentrifuge tubes
  • Hemoccult II Dispensapak Plus (Beckman Coulter, cat. no. 61130)
  • Coloview system (Karl Storz Veterinary Endoscopy)
  • Serum separator tubes: BD Microtainer tubes (BD Biosciences, cat. no. 365956)
  • Hitachi F‐4500 fluorescence spectrophotometer
  • Scalpel
  • Photographic equipment
  • 5‐ to 10‐ml syringes with 18‐G, 3‐in. straight 2.25‐mm ball needles (Braintree Scientific)
  • UltraTurax homogenizer (IKA works)
  • Refrigerated centrifuge
  • 96‐well plates
  • Human neutrophil MPO (Sigma, cat. no. M6908)
  • Toothpicks
  • 24‐well plates
  • 37°C, 5.0% CO 2 incubator
  • Additional reagents and equipment for administering inhaled anesthesia to mice (Isoflurane, anesthesia induction chamber, Parkland Scientific, cat. no. 93805107), fixing tissue in 10% PBS‐buffered formalin (Fisher, cat. no. SF100‐4, Chassaing et al., ), hematoxylin and eosin (H&E) staining (Thermo, cat. no. 7231 and 7111; Chassaing et al., ), immunohistochemistry (BrdU staining kit, Invitrogen, cat. no. 93‐3943), RNA extraction (Trizol, Invitrogen, cat. no. 15596‐026), myeloperoxidase assay (Sigma, cat. no. H9151, D3252, 216763, and M6908), and fecal biomarker assay (R&D Systems, cat. no. DY1857)
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Figures

Videos

Literature Cited

Literature Cited
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