An Air Pouch Mouse Model for Lipopolysaccharide-Induced Inflammatory Exudate Collection
Published: September 29, 2023
Abstract
Source: Lu, Q., et al. Detecting Migration and Infiltration of Neutrophils in Mice. J. Vis. Exp. (2020).
This video describes the procedure for generating an air pouch in the subcutaneous tissue of mice and collecting inflammatory exudates in response to lipopolysaccharide administration in vivo. This video also covers the analysis of the neutrophil infiltration in the air pouch.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Air pouch assay First air injection On day 0, fully anesthetize mice with 5% isoflurane for 3 min in a gas anesthesia chamber at a speed of 2 L/min, and maintain the anesthesia of each mouse in a single breathing unit with 2% isoflurane at a speed of 0.5 L/min. NOTE:…
Representative Results
Figure 1: Representative results of the air pouch assay. (A) Illustration of the air pouch assay. (B) Representative results of leukocyte subset infiltration in the air pouch assay. PBS: control; LPS: 1 µg/mL LPS. Data are presented as the mean ± SD.
Disclosures
The authors have nothing to disclose.
Materials
| 100% Ethanol | Beijing Chemical Works | ||
| 100% Methanol | Beijing Chemical Works | ||
| 15 mL Conical Polypropylene Centrifuge Tube | Falcon | Falcon | |
| 23 G x 1 1/4" Needle | BD | 305120 | |
| 26 G x 3/8" Needle | BD | 305110 | |
| 30 G x 1/2" Needle | BD | 305106 | |
| 5 mL Syringe | BD | Z683574 | |
| 50 mL Conical Polypropylene Centrifuge Tube | Falcon | 14-432-22 | |
| Gas Anesthesia System | ZS Dichuang | ZS-MV-IV | |
| Phosphate-Buffered Saline (PBS), 1× | Mix 90% ddH2O with 10% (v/v) 10×PBS, autoclaved | ||
| Phosphate-Buffered Saline (PBS), 10× | Dissolve 16 g NaCl, 0.4 g KCl, 2.88 g Na2HPO4·2H2O, 0.48 g KH2PO4 (anhydrous) in 200 mL ddH2O, adjust pH 7.4, autoclaved | ||
| Lipopolysaccharide (LPS) | Sigma | L3012 | |
| Wash Buffer in Air Pouch Assay | Dilute 0.5M EDTA to 10mM with HBSS |
Tags
Cite This Article
An Air Pouch Mouse Model for Lipopolysaccharide-Induced Inflammatory Exudate Collection. J. Vis. Exp. (Pending Publication), e21708, doi: (2023).