An Air Pouch Mouse Model for Lipopolysaccharide-Induced Inflammatory Exudate Collection

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.

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