Primary culture of tracheal aspirate-derived human airway basal stem cells

Patient-specific airway basal stem cells (BSCs) can be derived from tracheal aspirate (TA) samples from intubated patients, thus providing an invaluable lung stem cell derivation method that bypasses the need for lung tissue. The primary culture of BSCs provides the ideal model to study the function and differentiation of the conducting lung epithelium. This protocol outlines the specific steps for isolation, culture maintenance, passaging, freezing, thawing, differentiation, and immunofluorescence characterization of human TA-derived airway BSCs. For complete details on the use and execution of this protocol, please refer to Lu et al. (2021).