Abstract
A basic Bronchoalveolar lavage (BAL) procedure in mouse is described here. Cells and fluids obtained from BAL can be analyzed by Hema3-staining, immunostaining, Fluorescence-activated cell sorting (FACS), PCR, bicinchoninic acid protein assay, enzyme-linked immunosorbent assay (ELISA), luminex assays, etc., to examine the immune cells, pathogens, proteins such as cytokines/chemokines, and the expression levels of inflammation-related and other genes in the cells. This will help to understand the underlying mechanisms of these lung diseases and develop specific and effective drugs.
Keywords: Bronchoalveolar lavage, Lung cancer, Hema3-staining
Background
Bronchoalveolar lavage (BAL) is a simple and typical method commonly performed to diagnose pulmonary diseases including lung cancer (Daubeuf and Frossard, 2012). It is used to sample pulmonary components to determine the protein composition, immune cells and pathogens in the lung. Pulmonary chronic inflammation plays a critical role in lung cancer initiation and progression. To clarify the underlying mechanism of inflammation in lung tumorigenesis, a basic BAL protocol in mice is used in our laboratory to determine the pulmonary immune response (Qu et al., 2015; Zhou et al., 2015; Sun et al., 2016; Zhou et al., 2017).
Materials and Reagents
Equipment
Procedure
Data analysis
Murine BAL cells are retrieved and cytospun onto slides, which are Hema3-stained to determine the immune cells.
Notes
Although pelleted BAL cells from all four washes are combined to analyze the cells, you can combine the supernatant from only the first two washes (Procedure step 11) to analyze the BAL fluid components, as the first two washes contain a high concentration of these components.
Recipes
Acknowledgments
The authors thank the members in Xiao-Qu lab for the helpful discussion. This study was supported in part by the National Institute of Health (NIH)/National Cancer Institute (NCI) grants R01 CA172090, R21 CA175252, P50 CA090440, P30 CA047904, as well as the American Lung Association (ALA) Lung Cancer Discovery Award LCD 259111 and American Cancer Society (ACS) Fellowship Award PF-12081-01-TBG.
References
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