A Method for Culturing 3D Tumoroids of Lung Adenocarcinoma Cells at the Air–Liquid Interface

Three-dimensional (3D) tumor spheroids and tumoroids are among the most exploited cell culture methods in the lung cancer field, finding applications in the investigation of tumor growth and proliferation, invasion, and drug screening. However, 3D tumor spheroids and tumoroids cannot fully mimic the architecture of the human lung adenocarcinoma tissue and, in particular, the direct contact of the lung adenocarcinoma cells with the air, as they lack polarity. Our method allows to overcome this limitation by enabling to grow tumoroids of lung adenocarcinoma cells and healthy lung fibroblasts at the Air-Liquid Interface (ALI). This ensures straightforward access to both the apical and basal surface of the cancer cell culture, with several advantages in drug screening applications.