Isolation of highly pure alveolar epithelial type I and type II cells from rat lungs

• Published in: Laboratory investigation 2004-06, Vol.84 (6), p.727-735
• Main Authors: Chen, Jiwang, Chen, Zhongming, Narasaraju, Telugu, Jin, Nili, Liu, Lin
• Format: Article
• Language: English
• Subjects: Animals; Antibodies; Biological and medical sciences; Biotechnology; cell identification; Cell Separation - methods; Cell Survival; Epithelial Cells - cytology; Epithelial Cells - physiology; Fundamental and applied biological sciences. Psychology; hyperoxia; Hyperoxia - pathology; Hyperoxia - physiopathology; immunomagnetic separation; Immunomagnetic Separation - methods; Investigative techniques, diagnostic techniques (general aspects); Laboratory Medicine; Lung Injury; Male; Medical sciences; Medicine; Medicine & Public Health; Membrane Glycoproteins; Membrane Proteins - immunology; Microscopy, Electron; Pancreatic Elastase; Pathology; Pulmonary Alveoli - cytology; Pulmonary Alveoli - physiology; Pulmonary Surfactant-Associated Proteins - metabolism; Rats; Rats, Sprague-Dawley; Reproducibility of Results; research-article; type I and type II pneumocytes
• ISSN: 0023-6837; 1530-0307
• DOI: 10.1038/labinvest.3700095

There are no ideal cell lines available for alveolar epithelial type I and II cells (AEC I and II) at the present time. The current methods for isolating AEC I and II give limited purities. Here, we reported improved and reproducible methods for the isolation of highly pure AEC I and II from rat lungs. AEC I and II were released from lung tissues using different concentrations of elastase digestion. Macrophages and leukocytes were removed by rat IgG ‘panning' and anti-rat leukocyte common antigen antibodies. For AEC II isolation, polyclonal rabbit anti-T1α (an AEC I apical membrane protein) antibodies were used to remove AEC I contamination. For AEC I isolation, positive immunomagnetic selection by polyclonal anti-T1α antibodies was used. The purities of AEC I and II were 91±4 and 97±1%, respectively. The yield per rat was ∼2 × 106 for AEC I and ∼33 × 106 for AEC II. The viabilities of these cell preparations were more than 96%. The protocol for AEC II isolation is also suitable to obtain pure AEC II (93–95%) from hyperoxia-injured and recovering lungs. The purified AEC I and II can be used for gene expression profiling and functional studies. It also offers an important tool to the field of lung biology.