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Updated: Jan 13, 2026
Flow Cytometric Isolation of Primary Murine Type II Alveolar Epithelial Cells for Functional and Molecular Studies
Published on: December 26, 2012
Isolation of Alveolar Type II Epithelial Cells from Neonatal, Juvenile, and Adult Murine Lungs Adaptable to
Pia Grimpe1, Lars Melcher1, Lothar Gröbe2
1Infection Immunology Research Group, Institute of Medical Microbiology and Hospital Hygiene, Health Campus Immunology, Infectiology and Inflammation, Otto-von-Guericke University Magdeburg; Immune Regulation Research Group, Helmholtz Centre for Infection Research.
Abstract:
Alveolar type II epithelial cells (AECII) critically contribute to immune regulation in the lung and thus are key elements orchestrating pulmonary homeostasis. These functions render AECII particularly relevant in the context of infection with respiratory viruses, which target the lower airway epithelium and disrupt alveolar integrity. Preclinical mouse models are vital tools to elucidate the roles of AECII in the context of lung inflammation and/or infection. This protocol presents a detailed and reproducible pipeline for isolating viable and highly purified AECII from murine lungs at neonatal, juvenile, and adult stages. The methodology combines mechanical dissociation, enzymatic digestion, and subsequent fluorescence-activated cell sorting (FACS) to yield highly enriched AECII populations proven to be suitable for downstream molecular and functional analyses. For applications involving pathogenic viruses, the protocol includes an optional paraformaldehyde (PFA) fixation step, enabling safe handling under lower biosafety conditions without compromising RNA integrity. With optimized cell recovery across developmental stages, yielding up to 1.0 × 106 AECII cells per adult lung, this protocol supports preclinical studies of, e.g., viral pathogenesis, epithelial function, and regenerative capacity.
