Purpose

The incidence of acute lung injury (ALI) in severe trauma patients is 48% and the mortality rate following acute respiratory distress syndrome evolved from ALI is up to 68.5%. Alveolar epithelial type 1 cells (AEC1s) and type 2 cells (AEC2s) are the key cells in the repair of injured lungs as well as fetal lung development. Therefore, the purification and culture of AEC1s and AEC2s play an important role in the research of repair and regeneration of lung tissue.

Methods

Sprague-Dawley rats (3-4 weeks, 120-150 g) were purchase for experiment. Dispase and DNase I were jointly used to digest lung tissue to obtain a single-cell suspension, and then magnetic bead cell sorting was used to isolate AEC1s from the single-cell suspension by using AEC1s cell membrane marker protein T1a. Afterwards, alveolar epithelial cell membrane marker protein EpCAM was used to sort the remaining AEC2s. Cell purity was identified by immunofluorescence staining and flow cytometry.

Results

The purity of AEC1s and AEC2s was 88.3% ± 3.8% and 92.6% ± 2.7%. The cell growth was observed as follows: AEC1s stretched within the 12 h-16 h, but the cells proliferated slowly; while AEC2s began to stretch after 24 h and proliferated rapidly from the 2nd day and began to differentiate after 3 days.

Conclusion

AEC1s and AEC2s sorted by this method have high purity and good viability. Therefore, our method provides a new approach for the isolation and culture of AEC1s and AEC2s as well as a new strategy for the research of lung repair and regeneration.