Performing Bronchoalveolar Lavage in the Mouse

Abstract

Bronchoalveolar lavage (BAL) is a simple technique commonly used in humans to sample the contents of the epithelial lining fluid and determine the cellular and molecular composition of the pulmonary airways. In murine models, BAL makes it possible to sample immunological and inflammatory cell populations; it is indispensable for studying cell influx in disease models of the airways such as asthma and COPD. Cell counts can be combined with methods such as ELISA, immunoblot, immunohistochemistry, quantitative polymerase chain reaction, and HPLC to assess such inflammatory components as cytokines, growth factors, analytes, and receptors expressed at the cell membrane. Performing BAL in a reproducible manner is a hallmark of airway research in the mouse. Several procedures may be implemented. This unit describes a basic, rapid, inexpensive, and highly reproducible procedure to collect BAL fluid and cells that can be counted efficiently and reproducibly. Curr. Protoc. Mouse Biol. 2:167-175 © 2012 by John Wiley & Sons, Inc.

Atg7 Deficiency Intensifies Inflammasome Activation and Pyroptosis in Pseudomonas Sepsis

“…MH-S cells were obtained from American Type Culture Collection (ATCC, Manassas, VA). Bone marrow-derived macrophages (BMDM) were isolated from the femur of C57BL/6J wild-type (WT) mice and atg7 fl/fl mice after challenged with tamoxifen (29), and cultured in DMEM+10% FBS+10 ng/ml GM-CSF (R&D systems, Minneapolis, MN, without tamoxifen) (12). AMs were isolated by BAL and maintained in RPMI 1640 (Invitrogen, Grand Island, NY) supplemented with 1% penicillin streptomycin glutamine and 10% fetal bovine serum (FBS).…”

Cutaneous vaccination with coated microneedles prevents development of airway allergy

“…Goat anti mouse IgG, IgE, IgG1, and IgG2a antibodies labelled with horseradish peroxidase (Southern Biotech, Birmingham, Al, USA) were used as detection antibodies in ELISA. BAL fluid was collected as described previously [27]. In brief, the trachea region of a mouse was exposed by using a scissor, and then a 21-G catheter was placed into the trachea.…”

Fibroblasts secrete Slit2 to inhibit fibrocyte differentiation and fibrosis

“…injections of 2 μg of recombinant mouse Slit2 (R&D Systems) in 0.05 mL of PBS, or PBS alone, from day 1-14 after bleomycin aspiration (13 d total). Mice were euthanized 21 d after bleomycin aspiration, and bronchoalveolar lavage (BAL) cells were collected and counted, and cytospins were prepared as described previously (39,41,42,60). After BAL, lungs were inflated with and embedded in optimal cutting temperature (OCT) compound (VWR), frozen, and stored as described previously (23,39).…”