Isolation of Primary Alveolar Type II Cells
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Corresponding Organization : Tianjin International Joint Academy of Biomedicine
Other organizations : Chinese Academy of Medical Sciences & Peking Union Medical College, Peking Union Medical College Hospital, First Affiliated Hospital of Zhengzhou University
Variable analysis
- Lung digestion with dispase II (2 mg/ml) and deoxyribonuclease I (0.3 U/ml) at 37°C for 45 min
- Isolation and culture of primary AT2 cells
- Lavage of lungs with phosphate-buffered saline (PBS) containing 1% antibiotics
- Filtration of digested lung pieces through 100- and 40-μm cell strainers
- Removal of inflammatory cells using CD16/32- and CD45-coated dishes
- Removal of fibroblasts by further culturing the supernatant
- Seeding of cells in collagen I (BD)–precoated plates
- Culturing cells at 37°C with 5% CO2
- Positive control: Immunofluorescence staining using antibody against Prospc (Santa Cruz Biotechnology) to characterize AT2 cells
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