Open Access

Svenja NellingerORCiD, Simon HeineORCiD, Lothar Steeb, Petra KlugerORCiD

• In vitro alveolar models are an important tool study respiratory physiology, investigate lung diseases, develop and test new therapies, and simulate inhalation exposure to environmental pollutants or therapeutic agents. A defined cell culture medium is essential for maintaining consistent and reliable in vitro models. In this study a defined cell culture medium for advanced alveolar models based on epithelial A549 cell line and endothelial EaHy926 cell line is developed. Cellular survival was proven by quantification of LDH release. It was shown that co-culture of the cells enhances cellular survival compared to the endothelial monoculture. The formation of a homogeneous monolayer on laminin coated membranes was confirmed by actin staining. Further, the expression of cell type specific proteins (endothelial: CD31, VE-cadherin; epithelial: ZO1 and E-cadherin) was shown. Overall, we demonstrated a completely defined cell culture medium that can be used for the setup of an advanced in vitro alveolar model for biomedical application like drug development or breath gas analysis.