Long-Term Culture of Distal Airway Epithelial Cells Allows Differentiation Towards Alveolar Epithelial Cells Suited for Influenza Virus Studies

Imai-Matsushima, Aki; Martin-Sancho, Laura; Karlas, Alexander; Imai, Seiichiro; Zoranovic, Tamara; Hocke, Andreas C.; Mollenkopf, Hans-Joachim; Berger, Hilmar; Meyer, Thomas F.

Immunofluorescence labeling showed that the majority of human DAECs expressed both cytokeratin 5 (CK5) and the N-terminal truncated form of P63 (ΔN-P63), considered the main markers of regenerating cells in the mouse lung after severe injury ( Fig. 2A , B) [16] . Flow cytometry analyses further revealed that 80.6% of E-cadherin + DAECs were double positive for α6 and β4 integrins (Fig. 2C) , markers of regenerating cells in mouse lung [25] . Regenerating cells are negative for markers of AEII cells and airway club cells -the main mature epithelial cells with proliferation potential in the human distal lung. As expected, DAECs were negative for the AEII cell marker pro-surfactant protein C (proSP-C) at passage 4 ( Fig. 2D ), although some were positive at earlier passages (Suppl. Fig. 1 ). DAECs were also negative for the airway club cell marker secretoglobin1A1 (SCGB1A1) (Fig. 2E ).