Name: Murine Distal Colostomy, A Novel Model of Diversion Colitis in C57BL/6 Mice (Video) | JoVE Journal
Uploaded: 2018-07-12T13:30:00.000+00:00
Duration: 8 min 20 s
Description: Learn a murine distal colostomy model for studying diversion colitis in C57BL/6 mice. This technique enables immune-microbiome interaction research without requiring genetically modified animals.

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5 minutes

All methods described here have been approved by the veterinary government authority (Landesamt f&#252;r Landwirtschaft, Lebensmittelsicherheit und Fischerei Mecklenburg-Vorpommern, (LALLF M-V)).
1. Preoperative Care and Preparation of the Animal
<ol>
	<li>Upon arrival in the animal facility, divide animals (C57Bl/6) into groups of similar size, cage each group together, and keep groups constant throughout the experiments. 
	NOTE: Use animals of the same sex. The results described were obtained with male mice. 
	NOTE: If male animals are used, cage groups together starting when they are 7 weeks of age so that a hierarchy can be established, thus minimizing the risk of aggressive behavior during experiments.</li>
	<li>At least one week before surgery, switch to a high energy (&#62;14 MJ/kg) and high protein (&#62;20%) feed containing all the essential trace elements and vitamins (for details, see list of materials). 
	NOTE: Assure all mice weigh at least 25 g when surgery is performed.</li>
	<li>Induce anaesthesia and analgesia by intraperitoneal injection of ketamine (87 mg/kg i.p.) and xylazine hydrochloride (13 mg/kg i.p.).&#160;Wait until the mouse tolerates mechanical stimulation, e.g. toe pitch, without motor response.</li>
	<li>Secure the narcotized mouse with tapes in a supine position on a heat underlay positioned at the operation desk, guaranteeing stable positioning during operation and avoiding an overwhelming loss of body heat. 
	NOTE: The heat underlay should have a surface temperature of 36 &#176;C to 40 &#176;C; the operating room temperature should be 21 &#176;C.</li>
</ol>
2. Distal Colostomy Operation
<ol>
	<li>Shave the abdominal hair. Before starting surgery, disinfect the operation field three times using alcohol 70% and an iodophor. Drape the operation field to guarantee for aseptic conditions.</li>
	<li>Perform a 15-mm median laparotomy by incising the abdominal muscles and the peritoneum along the linea alba, thus minimizing blood loss.</li>
	<li>Use two DeBakey atraumatic forceps, carefully pull the cecum, terminal ileum, and ascending and transverse colon from the peritoneal cavity. 
	NOTE: Be careful to strictly limit mechanical manipulation of the intestine to prevent injury to mesenteric structures.</li>
	<li>Identify the cecal pole, the ascending colon, and small intestine (Figures 1a and 1b). 
	NOTE: Correct identification of the ascending colon is fundamental for correct placement of the colostomy. In cases where the anatomy of the ileocecal region is ambiguous, the presence of Peyer&#39;s patches identifies the small intestine, and the presence of formed stool characterizes the colon.</li>
	<li>Use a ruler to determine the position of the future colostomy. It should be placed 20 mm distal to the ileocecal valve for a distal colostomy.</li>
	<li>Make a second 3-mm incision in the abdominal wall in the upper right quadrant. Pull the previously identified colon segment through this incision to form a loop, being careful not to distort the loop.</li>
	<li>Carefully pass a 22-gauge flexible i.v. cannula through the mesocolon. Take care not to damage mesenteric vascular structures.</li>
	<li>Return the intestine to the peritoneal cavity.</li>
	<li>Fix both ends of the flexible tube to the skin using simple stitches and a resorbable suture (e.g., polyglactin 910 or polyfil 4-0 1/2c).</li>
	<li>Before closing the laparotomy, perform fluid resuscitation using an intraperitoneal injection of 0.5 mL 0.9% saline.</li>
	<li>Close the peritoneum and the muscle layer with a continuous suture using a resorbable suture (e.g., polyglactin 910 or polyfil 4-0 1/2c). Close the skin with a continuous suture using a resorbable suture (e.g., polyglactin 910 or polyfil 4-0 1/2c).</li>
	<li>Open the exteriorized colon loop by performing a subtotal transection using a fine scissor. Avoid all injury to the mesentery. Do not transect the colon completely.</li>
	<li>Fix each colostomy opening using three single full-thickness stitches to the peritoneum and skin using a monofil, absorbable suture (e.g., polydioxanone or monofil 6-0 3/8s). The afferent loop, which is a functional end-colostomy, and the efferent loop, which is a mucous fistula, are clearly separated at this point (Figure 1c). 
	NOTE: Operation time should be less than 20 minutes to limit fluid and thermal losses.</li>
	<li>After finishing surgery, disinfect instruments using an aldehyde-free disinfection solution in an ultrasonic bath according to the instructions of the manufacturer.</li>
</ol>
3. Sham Operation (Colotomy)
<ol>
	<li>Perform steps 1.1. through 2.4.</li>
	<li>Use a ruler to determine the future colotomy position. The colotomy should be positioned the same distance from the ileocecal valve as the colostomy in the experimental group.</li>
	<li>Open the colon at least two-thirds its circumference using fine scissors.</li>
	<li>Close the colotomy with a single layer, full-thickness interrupted suture using a monofil, absorbable suture (e.g., polydioxanone, monofil 6-0 3/8s). 
	NOTE: Operation time should be less than 20 minutes for an experimented surgeon, limiting fluid and thermal losses.</li>
	<li>Perform steps 2.10. , 2.11. and 2.14.</li>
</ol>
4. Postoperative Care
<ol>
	<li>Return animals to their cages. Provide a well-tempered atmosphere of 37 &#176;C (e.g., with an infrared lamp) until mice are fully awake. Then, keep mice in a temperature- and humidity-regulated environment (21 &#176;C; 30% &#177; 10% relative humidity). Allow free access to food and drinking water. To facilitate fluid uptake, provide additional water-soaked animal feed.</li>
	<li>Start postoperative analgesia by injecting 0.1 mg/kg body weight buprenorphine s.c. when animals show response to mechanical stimulation. Be careful to avoid respiratory depression.</li>
	<li>Supplement drinking water with 1 mg/mL tramadol for continued analgesia during the first postoperative week.</li>
	<li>To compensate for decreased fluid intake due to reduced mobility, supply a Solid Drink Pad in the cage during the first postoperative week.</li>
	<li>Weigh the animals and score animal behavior daily during the first week, every second day during remainder of the first month, and every third day during the second month using the disease severity score described in Kleinwort et al.6.</li>
</ol>

<ol><li>Glotzer, D. J., Glick, M. E., Goldman, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Proctitis+and+colitis+following+diversion+of+the+fecal+stream%5BTitle%5D)+AND+%22Gastroenterology%22%5BJournal%5D)">Proctitis and colitis following diversion of the fecal stream.</a> Gastroenterology. 80, 438-441 (1981).</li>
<li>Morson, B., Dawson, I. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aMorson%2C+B+%22Gastrointestinal+Pathology%22">Gastrointestinal Pathology</a>. , Blackwell Scientific Publications. London. (1972).</li>
<li>Whelan, R. L., Abramson, D., Kim, D. S., Hashmi, H. F. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/8153859">Diversion colitis. A prospective study.</a> Surg Endosc. 8, 19-24 (1994).</li>
<li>Haas, P. A., Fox, T. A., Szilagy, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Endoscopic+examination+of+the+colon+and+rectum+distal+to+a+colostomy%5BTitle%5D)+AND+%22Am+J+Gastroenterol%22%5BJournal%5D)">Endoscopic examination of the colon and rectum distal to a colostomy.</a> Am J Gastroenterol. 85, 850-854 (1990).</li>
<li>Valatas, V., Bamias, G., Kolios, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Experimental+colitis+models%3A+Insights+into+the+pathogenesis+of+inflammatory+bowel+disease+and+translational+issues%5BTitle%5D)+AND+%22Eur+J+Pharmacol%22%5BJournal%5D)">Experimental colitis models: Insights into the pathogenesis of inflammatory bowel disease and translational issues.</a> Eur J Pharmacol. 759, 253-264 (2015).</li>
<li>Kleinwort, A., Döring, P., Hackbarth, C., Heidecke, C. D., Schulze, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Deviation+of+the+Fecal+Stream+in+Colonic+Bowel+Segments+Results+in+Increased+Numbers+of+Isolated+Lymphoid+Follicles+in+the+Submucosal+Compartment+in+a+Novel+Murine+Model+of+Diversion+Colitis%5BTitle%5D)+AND+%22Biomed+Res+Int%22%5BJournal%5D)">Deviation of the Fecal Stream in Colonic Bowel Segments Results in Increased Numbers of Isolated Lymphoid Follicles in the Submucosal Compartment in a Novel Murine Model of Diversion Colitis.</a> Biomed Res Int. 2017, 13(2017).</li>
<li>Tangjarukij, C., Navasumrit, P., Zelikoff, J. T., Ruchirawat, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+effects+of+pyridoxine+deficiency+and+supplementation+on+hematological+profiles%2C+lymphocyte+function%2C+and+hepatic+cytochrome+P450+in+B6C3F1+mice%5BTitle%5D)+AND+%22J+Immunotoxicol%22%5BJournal%5D)">The effects of pyridoxine deficiency and supplementation on hematological profiles, lymphocyte function, and hepatic cytochrome P450 in B6C3F1 mice.</a> J Immunotoxicol. 6, 147-160 (2009).</li>
<li>Soave, O., Brand, C. D. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/1954740">Coprophagy in animals: a review.</a> Cornell Vet. 81, 357-364 (1991).</li>
<li>Ebino, K. Y., Yoshinaga, K., Suwa, T., Kuwabara, Y., Takahashi, K. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effects+of+prevention+of+coprophagy+on+pregnant+mice--is+coprophagy+beneficial+on+a+balanced+diet%5BTitle%5D)+AND+%22Jikken+Dobutsu%22%5BJournal%5D)">Effects of prevention of coprophagy on pregnant mice--is coprophagy beneficial on a balanced diet.</a> Jikken Dobutsu. 38, 245-252 (1989).</li>
<li>Babickova, J., Tothova, L., Lengyelova, E., Bartonova, A., Hodosy, J., Gardlik, R., Celec, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Sex+Differences+in+Experimentally+Induced+Colitis+in+Mice%3A+a+Role+for+Estrogens%5BTitle%5D)+AND+%22Inflammation%22%5BJournal%5D)">Sex Differences in Experimentally Induced Colitis in Mice: a Role for Estrogens.</a> Inflammation. 38, 1996-2006 (2015).</li>
<li>Lee, S. M., Kim, N., Son, H. J., Park, J. H., Nam, R. H., Ham, M. H., Choi, D., Sohn, S. H., Shin, E., Hwang, Y. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+Effect+of+Sex+on+the+Azoxymethane%2FDextran+Sulfate+Sodium-treated+Mice+Model+of+Colon+Cancer%5BTitle%5D)+AND+%22J+Cancer+Prev%22%5BJournal%5D)">The Effect of Sex on the Azoxymethane/Dextran Sulfate Sodium-treated Mice Model of Colon Cancer.</a> J Cancer Prev. 21, 271-278 (2016).</li>
<li>Angele, M. K., Pratschke, S., Hubbard, W. J., Chaudry, I. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Gender+differences+in+sepsis%3A+cardiovascular+and+immunological+aspects%5BTitle%5D)+AND+%22Virulence%22%5BJournal%5D)">Gender differences in sepsis: cardiovascular and immunological aspects.</a> Virulence. 5, 12-19 (2013).</li>
<li>Brown, R. Z. <a target="_blank" href="http://onlinelibrary.wiley.com/doi/10.2307/1943592/abstract">Social Behaviour, reproduction, and population changes in the house mouse (Mus musculus L).</a> Eccological Monographs. 23, 788-795 (1953).</li>
<li>Poole, T. B., Morgan, H. D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Differences+in+aggressive+behaviour+betweeen+male+mice+%28Mus+musculus+L.%29+in+colonies+of+different+sizes%5BTitle%5D)+AND+%22Animal+Behaviour%22%5BJournal%5D)">Differences in aggressive behaviour betweeen male mice (Mus musculus L.) in colonies of different sizes.</a> Animal Behaviour. 21, 788-795 (1973).</li>
<li>Pasquarelli, N., Voehringer, P., Henke, J., Ferger, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effect+of+a+change+in+housing+conditions+on+body+weight%2C+behavior+and+brain+neurotransmitters+in+male+C57BL%2F6J+mice%5BTitle%5D)+AND+%22Behav+Brain+Res%22%5BJournal%5D)">Effect of a change in housing conditions on body weight, behavior and brain neurotransmitters in male C57BL/6J mice.</a> Behav Brain Res. 333, 35-42 (2017).</li>
<li>Langgartner, D., Foertsch, S., Fuchsl, A. M., Reber, S. O. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/27788633">Light and water are not simple conditions: fine tuning of animal housing in male C57BL/6 mice.</a> Stress. 20, 10-18 (2017).</li>
<li>Nicholson, A., Malcolm, R. D., Russ, P. L., Cough, K., Touma, C., Palme, R., Wiles, M. V. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/19930822">The response of C57BL/6J and BALB/cJ mice to increased housing density.</a> J Am Assoc Lab Anim Sci. 48, 740-753 (2009).</li>
<li>Buchler, G., Wos-Oxley, M. L., Smoczek, A., Zschemisch, N. H., Neumann, D., Pieper, D. H., Hedrich, H. J., Bleich, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Strain-specific+colitis+susceptibility+in+IL10-deficient+mice+depends+on+complex+gut+microbiota-host+interactions%5BTitle%5D)+AND+%22Inflamm+Bowel+Dis%22%5BJournal%5D)">Strain-specific colitis susceptibility in IL10-deficient mice depends on complex gut microbiota-host interactions.</a> Inflamm Bowel Dis. 18, 943-954 (2012).</li>
<li>Nakanishi, M., Tazawa, H., Tsuchiya, N., Sugimura, T., Tanaka, T., Nakagama, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mouse+strain+differences+in+inflammatory+responses+of+colonic+mucosa+induced+by+dextran+sulfate+sodium+cause+differential+susceptibility+to+PhIP-induced+large+bowel+carcinogenesis%5BTitle%5D)+AND+%22Cancer+Sci%22%5BJournal%5D)">Mouse strain differences in inflammatory responses of colonic mucosa induced by dextran sulfate sodium cause differential susceptibility to PhIP-induced large bowel carcinogenesis.</a> Cancer Sci. 98, 1157-1163 (2007).</li>
<li>Mahler, M., Bristol, I. J., Leiter, E. H., Workman, A. E., Birkenmeier, E. H., Elson, C. O., Sundberg, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Differential+susceptibility+of+inbred+mouse+strains+to+dextran+sulfate+sodium-induced+colitis%5BTitle%5D)+AND+%22Am+J+Physiol%22%5BJournal%5D)">Differential susceptibility of inbred mouse strains to dextran sulfate sodium-induced colitis.</a> Am J Physiol. 274, 544-551 (1998).</li>
<li>Mekada, K., Abe, K., Murakami, A., Nakamura, S., Nakata, H., Moriwaki, K., Obata, Y., Yoshiki, A. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/19448337">Genetic differences among C57BL/6 substrains.</a> Exp Anim. 58, 141-149 (2009).</li>
<li>Sellers, R. S., Clifford, C. B., Treuting, P. M., Brayton, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Immunological+variation+between+inbred+laboratory+mouse+strains%3A+points+to+consider+in+phenotyping+genetically+immunomodified+mice%5BTitle%5D)+AND+%22Vet+Pathol%22%5BJournal%5D)">Immunological variation between inbred laboratory mouse strains: points to consider in phenotyping genetically immunomodified mice.</a> Vet Pathol. 49, 32-43 (2012).</li>
<li>Mills, C. D., Kincaid, K., Alt, J. M., Heilman, M. J., Hill, A. M. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/10843666">M-1/M-2 macrophages and the Th1/Th2 paradigm.</a> J Immunol. 164, 6166-6173 (2000).</li>
</ol>

All authors declare that they have no competing interests.

The murine model of DC presented in this protocol reliably reproduces the histopathological features of human DC (e.g., de novo development of lymphoid follicles in the submucosa of inflamed bowel segments, crypt shortening, and reductions in goblet cell numbers). Aside from this advantage, this model is induced by a very similar triggering factor and presents with a clinical course of moderate to mild severity as is the case in most affected humans.
To obtain reproducible re...

In recent years, a substantial number of non-infectious colitis entities different from classical inflammatory bowel diseases (IBDs; i.e., Crohn&#39;s disease or colitis ulcerosa) have been clinically and histopathologically characterized in humans. The pathophysiological mechanisms leading to the development of these colitis forms are not completely understood partially because appropriate animal models are scarce. Diversion colitis is one of these recently described entities. Although the term was coined in 1980 by Glotzer1, the initial description of a similar phenotype was given in 1972 by Morson2. The disease develops in 50% to 91% of patients with diverting enterostomy, and its clinical intensity varies3,4. Given the annual incidence of around 120,000 colostomy patients in the United States of America, this disease entity constitutes an important health problem.
The overall goal of developing this protocol was to provide a murine DC model that relies on a colitis trigger similar to that seen in human DC and that reproduces the primary histopathological features of the human disease. In contrast to other murine colitis models, colitis induction in our model does not require genetically modified animals (e.g., IL-7 transgenic mice, N-cadherin dominant negative mice, or TGF&#946;-/- mice), the application of chemically irritating substances (e.g., dextran sulfate sodium (DSS)-induced colitis, or trinitrobenzene sulfonic acid (TNBS)-induced colitis), or the transfer of specific cell populations in immune deficient mice (as in the CD45RBhigh transfer model of colitis) (for a review, see5). In contrast to other models, the intact immune system of our DC model allows assessment of the immunological mechanism involved in DC development. The limitation of mucosal inflammation to the excluded bowel segment allows assessment of its repercussion on mucosal immunity in other parts of the gastrointestinal tract, on the immune homeostasis in other immune compartments of the intestinal tract (e.g., Peyer&#39;s patches and mesenteriale lymph nodes), and on the immune homeostasis of the entire organism. Finally, our model constitutes an appropriate tool for investigating the mechanisms controlling local inflammatory stimuli originating from changes in the normal local environment for both the local microbiome as well as alimentary antigens.

<table><tbody><tr><td>&lt;em&gt;Operation material&lt;/em&gt;</td><td></td><td></td><td></td></tr><tr><td>heat underlay, 6 watt</td><td>ThermoLux, Witte + Sutor GmbH</td><td>461265</td><td></td></tr><tr><td>Ethanol 70% (with methylethylketon)</td><td>Pharmacie of the University Hospital Greifswald</td><td></td><td></td></tr><tr><td>Wooden applicators, small cotton head onesided, wooden stick, 150 mm length</td><td>Centramed GmbH, Germany</td><td>8308370</td><td>for disinfection of operation field</td></tr><tr><td>Scissor</td><td>Aesculap (BRAUN)</td><td>BC064R</td><td></td></tr><tr><td>Atraumatic DeBakey forceps</td><td>Aesculap (BRAUN)</td><td>OC021R</td><td></td></tr><tr><td>Needle holder</td><td>Aesculap (BRAUN)</td><td>BM012R</td><td></td></tr><tr><td>Vasofix Safety &lt;em&gt;i.v.&lt;/em&gt; cannula 22G</td><td>Braun Melsungen AG, Germany</td><td>4268091S-01</td><td></td></tr><tr><td>VICRYL Plus 4-0 violet braided; V-5 17 m 1/2c; 70 cm</td><td>ETHICON, Inc.</td><td>VCP994H</td><td></td></tr><tr><td>PDS II 6-0 monofil; V-18 13 mm 3/8c; 70 cm</td><td>ETHICON, Inc.</td><td>Z991H</td><td></td></tr><tr><td>BD Plastipak 1 mL (Syringes with needle with sterile interior)</td><td>BD Medical</td><td>305501</td><td></td></tr><tr><td>Triacid-N (N-Dodecylpropan-1,3-diamin)</td><td>ANTISEPTICA, Germany</td><td>18824-01</td><td>disinfection of surgical instruments in ultrasonic bath</td></tr><tr><td>&lt;em&gt;Medications&lt;/em&gt;</td><td></td><td></td><td></td></tr><tr><td>ketamine 10%, 100 mg/mL (ketamine hydrochloride)</td><td>selectavet, Dr. Otto Fischer GmbH</td><td>9089.01.00</td><td>87 mg/kg &lt;em&gt;i.p.&lt;/em&gt;</td></tr><tr><td>Xylasel, 20 mg/mL (xylazine hydrochloride)</td><td>selectavet, Dr. Otto Fischer GmbH</td><td>400300.00.00</td><td>13 mg/kg &lt;em&gt;i.p.&lt;/em&gt;</td></tr><tr><td>NaCl 0.9%</td><td>Braun Melsungen AG, Germany</td><td>6697366.00.00</td><td></td></tr><tr><td>Buprenovet 0.3 mg/mL (buprenorphine)</td><td>Bayer, Germany</td><td>PZN: 01498870</td><td>0.1 mg/kg &lt;em&gt;s.c.&lt;/em&gt;</td></tr><tr><td>Tramal Drops, 100 mg/mL (tramadol hydrochloride)</td><td>Gr&amp;uuml;nenthal GmbH, Germany</td><td>10116838</td><td>1 mg/mL drinking water</td></tr><tr><td>Ceftriaxon-saar 2 g (ceftriaxone)&amp;nbsp;</td><td>Cephasaar GmbH, Germany</td><td>PZN: 08844252</td><td>25 mg/kg body weight &lt;em&gt;i.p.&lt;/em&gt;</td></tr><tr><td>metronidazole 5 mg/mL</td><td>Braun Melsungen AG, Germany</td><td>PZN: 05543515</td><td>12.5 mg/kg body weight &lt;em&gt;i.p.&lt;/em&gt;</td></tr><tr><td>&lt;em&gt;Food&lt;/em&gt;</td><td></td><td></td><td></td></tr><tr><td>ssniff M-Z Ereich</td><td>ssniff, Germany</td><td>&amp;nbsp;V1184-3</td><td></td></tr><tr><td>Solid Drink Dehyprev Vit BIO, pouches</td><td>TripleATrading, the Netherlands</td><td>SDSHPV-75</td><td></td></tr><tr><td>Equipment</td><td></td><td></td><td></td></tr><tr><td>Nikon Eclipse Ci-L</td><td>Nikon Instruments Europe BV, Germany</td><td></td><td>light microscopy</td></tr><tr><td>VetScan HM 5&amp;nbsp;</td><td>Abaxis, USA</td><td>770-9000</td><td>Veterinary hematology analyzer of 50 &amp;micro;l venous EDTA-blood</td></tr><tr><td>Bandelin SONOREX (Ultrasonic bath)</td><td>Bandelin electronics, Germany</td><td>RK 100 H</td><td>disinfection of surgical instruments</td></tr><tr><td>&lt;em&gt;Software&lt;/em&gt;</td><td></td><td></td><td></td></tr><tr><td>NIS-Element BR4 software</td><td>Nikon Instruments Europe BV, Germany</td><td></td><td></td></tr><tr><td>GraphPad Prism Version 6</td><td>GraphPad Software, Inc.</td><td></td><td></td></tr></tbody></table>

murine distal colostomy, a novel model of diversion colitis in c57bl/6 mice

Murine Distal Colostomy, A Novel Model of Diversion Colitis in C57BL/6 Mice

Diversion colitis (DC) is a frequent clinical condition occurring in patients with bowel segments excluded from the fecal stream as a result of a diverting enterostomy. The etiology of this disease remains ill-defined but appears to differ from that of classical inflammatory bowel diseases such as Crohn's disease and ulcerative colitis. Research aimed to decipher the pathophysiological mechanisms leading to the development of this disease has been severely hampered by the lack of an appropriate murine model. This protocol generates a murine model of DC that facilitates the study of the immune system's role and its interaction with the microbiome in the development of DC. In this model using C57BL/6 animals, distal parts of the colon are excluded from the fecal stream by creating a distal colostomy, triggering the development of mild to moderate inflammation in the excluded bowel segments and reproducing the hallmark lesions of human DC with a moderate systemic inflammatory response. In contrast to the rat model, a large number of genetically-modified murine models on the C57BL/6 background are available. The combination of these animals with our model allows the potential roles of individual cytokines, chemokines, or receptors of bioactive molecules (e.g., interleukin (IL)-17; IL-10, chemokine CXCL13, chemokine receptors CXCR5 and CCR7, and the sphingosine-1-phosphate receptor 4) to be assessed in the pathogenesis of DC. The availability of congenic mouse strains on the C57BL/6 background largely facilitates transfer experiments to establish the roles of distinct cell types involved in the etiology of DC. Finally, the model offers the opportunity to assess the influences of local interventions (e.g., modification of the local microbiome or local anti-inflammatory therapy) on mucosal immunity in affected and non-affected bowel segments and the on systemic immune homeostasis.

Surgery is well tolerated both in the experimental (colostomy) and sham (colotomy) groups. Perioperative mortality should not exceed 10% when surgery and perioperative management is correctly performed. In the first postoperative week, significant weight loss is seen in both the experimental and sham groups. Animals of the sham group attain their weight nadir usually toward the fourth postoperative day, but it occurs one day later in the experimental group. Weight loss is more pronounced ...

Murine distal colostomy provides a murine model for human diversion colitis, a predominantly lymphocytic colitis in colon segment excluded from the fecal stream.

Diversion colitis (DC) is a frequent clinical condition occurring in patients with bowel segments excluded from the fecal stream as a result of a ...

murine-distal-colostomy-novel-model-diversion-colitis-c57bl6

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In this video, we describe a surgical procedure to generate mouse proximal colon loop or pcLoop segment followed by intraluminal injection of fluorescent markers into the segment to study the intestinal epithelial permeability.

Generation of Mouse Proximal Colon Loop Model: A Surgical Procedure to Generate Proximal Colon Loop and Measure Intestinal Permeability by Detection of Fluorescent Markers in Serum Following Intraluminal Injection

This video demonstrates a technique to generate a chemical-induced murine model of colitis — an inflammatory bowel disease. The mouse is allowed to consume a dextran sodium sulfate (DSS) solution for an adequate duration. Upon consumption, DSS damages the mucus and epithelial layer barrier in the colon, allowing gut microbes to enter the underlying tissue — where they are sensed by the resident immune cells to mediate an exaggerated immune response, resulting in inflammation.

A Technique to Generate a Murine Model of Dextran Sulfate Sodium-Induced Colitis

Citrobacter rodentium infection provides a valuable model to study enteric bacterial infections as well as host immune responses and colitis in mice. This protocol outlines the measurement of barrier integrity, pathogen load and histological damage allowing for the thorough characterization of pathogen and host contributions to murine infectious colitis.

The Citrobacter rodentium Mouse Model: Studying Pathogen and Host Contributions to Infectious Colitis

Over the past few years, new generation endoscopes have emerged as important diagnostic research aids for evaluating murine colitis and colorectal tumors. We present herein a detailed protocol for endoscopic assessment of inflammation and colorectal tumors in mice, as well as a novel scoring system that uses decimal identifiers to document the endoscopic severity of colitis and colorectal tumors.

Flexible Colonoscopy in Mice to Evaluate the Severity of Colitis and Colorectal Tumors Using a Validated Endoscopic Scoring System

A protocol for the establishment of a genetically engineered mouse model of colorectal cancer by segmental adeno-cre infection and its surveillance via high-resolution colonoscopy is presented.

A Genetically Engineered Mouse Model of Sporadic Colorectal Cancer

Intestinal obstructions are a partial or complete blockage of the intestine that can cause severe abdominal pain, nausea, vomiting, and preventing the passage of stool. This procedure for creating intestinal partial obsructions in mice is reliable in studying the mechanisms underlying pathological cell growth and death in the intestine.

A Mouse Model of Intestinal Partial Obstruction

Anastomotic leak or breakdown after surgery is a major cause of postoperative morbidity and mortality. Our surgery for creating a colonic anastomosis is a reliable and reproducible method for studying the healing mechanisms of anastomoses.

Creation of Colonic Anastomosis in Mice

The presented protocol describes a facile surgical removal of the appendix (caecal patch) in a mouse followed by the induction of inflammatory bowel disease-associated colorectal cancer. This murine appendectomy model enables investigation of the biological role of the appendix in the pathogenesis of human gastrointestinal disease.

Murine Appendectomy Model of Chronic Colitis Associated Colorectal Cancer by Precise Localization of Caecal Patch

We describe a murine model of postoperative ileus generated via intestinal manipulation. Gastrointestinal transit function, pathologic changes, and immune cell activation were assessed 24 h after surgery.

Postoperative Ileus Murine Model

Learn a murine distal colostomy model for studying diversion colitis in C57BL/6 mice. This technique enables immune-microbiome interaction research without requiring genetically modified animals.

Murine Distal Colostomy, A Novel Model of Diversion Colitis in C57BL/6 Mice

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