Figure 1.

Preparation of single cell suspensions from neonatal and adult murine lungs for isolation of APCs. (A) To prepare single cell suspensions, neonatal and adult mice were euthanized and the lungs were exposed by dissecting through the thorax by raising the sternum to avoid any injury to the lung, perfused through right ventricle by adding ice cold PBS (containing 2 mM EDTA) and lung lobes were carefully isolated and were manually minced into small pieces in Petri dishes. Minced pieces were digested and digested lung tissue was subsequently passed through a cell strainer. For multicolor immunophenotyping and in vitro stimulation assay, lung APCs were enriched using CD11c- based magnetic beads for use in in vitro stimulation assays. (B) List of known pan- and precursor- markers used in combination to identify heterogeneous populations of lung APCs. (C) Overview of the lung APCs studied in this study.