This video demonstrates the generation of an ileal loop model to study intestinal permeability in a mouse model. This method provides a live model to understand the epithelial barrier function and consequent immune response.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Generation of the ileal loop Skin preparation: Scrub fur of the abdominal midline with alcohol swabs or gauze sponge soaked with 70% Ethanol. Do not wet a wide area of fur with alcohol to prevent hypothermia. Using scissors, perform a midline laparotomy. Make a vertical incision in the middle of the abdomen (a…
Representative Results
Figure 1: The ileal loop model. (A) Schematic overview of the ileal loop model. Median laparotomy is performed on mice under anesthesia and placed on a temperature-controlled surgery board. (B) Exteriorization of the caecum (*), ileum and mesentery. Two adequate sites for ligation are identified (1,2). (C) Isolate a segment of 4 cm length: the first ligature (1) is placed close to the ileo-caecal…
Disclosures
The authors have nothing to disclose.
Materials
Equipment and Material
BD Alcohol Swabs
BD
326895
BD PrecisionGlide Needle, 25G X 5/8"
BD
305122
BD PrecisionGlide Needle, 30G X 1/2"
BD
305106
BD 1ml Tuberculin Syringe Without Needle
BD
309659
15ml Centrifuge Tube
Corning
14-959-53A
Corning 96-Well Solid Black Polystyrene Microplate
FisherScientific
07-200-592
Corning Non-treated Culture Dish, 10cm
MilliporeSigma
CLS430588
Cotton Tip Applicator (cotton swab), 6", sterile
FisherScientific
25806 2WC
Dynarex Cotton Filled Gauze Sponges, Non-Sterile, 2" x 2"
Medex
3249-1
EZ-7000 anesthesia vaporizer (Classic System, including heating units)
E-Z Systems
EZ-7000
Moria Fine Scissors
FST
14370-22
Puralube Vet Ointment, Sterile Ocular Lubricant
Dechra
12920060
Ring Forceps (blunt tissue forceps)
FST
11103-09
Roboz Surgical 4-0 Silk Black Braided, 100 YD
FisherScientific
NC9452680
Semken Forceps (anatomical forceps)
FST
1108-13
Sofsilk Nonabsorbable Coated Black Suture Braided Silk Size 3-0, 18", Needle 19mm length 3/8 circle reverse cutting
HenrySchein
SS694
Student Fine Forceps, Angled
FST
91110-10
10ml Syringe PP/PE without needle
Millipore Sigma
Z248029
Yellow Feeding Tubes for Rodents 20G x 30 mm
Instech
FTP-20-30
Solutions and Buffers
Hanks' Balanced Salt Solution
Corning
21-023-CV
Phosphate-Buffered Saline without Calcium and Magnesium