Human lung microvascular endothelial cells

Our LOMM contains NBECs collected from 5 healthy donors (Supplementary Table 1). We generated the lung models as we described previously in [8 (link)] by culturing five × 10⁵ cells NBECs of each donor (5 donors × 3 replicates for each donor = 15 experiments) on the top side of transwell® polyester membrane cell culture inserts and human endothelial cells (2 × 10⁵ cells, Human Lung Microvascular Endothelial Cells, Lonza, Walkersville, MD) on the bottom side at the air–liquid interface (ALI). The LOMM models were split into a control (unchallenged group), challenged LOMM (challenged with MAB microparticle), and challenged LOMM + α-MSH (challenged with MAB microparticle and treated with 5 μM α-MSH (Bachem Americas, Inc, CA USA) every 24 h. In total, five LOMM were included in each group. In addition, one group received α-MSH (not challenged with MAB microparticles) and was another control, only for western blotting analysis of type I IFN proteins (MX1, OSA1, and ISG15). The cell culture media of all wells were collected after 48 h for measurements.

The following cell lines and cell types were propagated using established methods or according to the manufacturer’s instructions. A549, H441 cell lines, Human Bronchial Epithelial Cells (ATCC, VA), Human Lung Microvascular Endothelial Cells (Lonza, MD), T7 cells (mouse type II alveolar epithelial cell line; Sigma-Aldrich, MO), C22 cells (mouse club cell line; Sigma-Aldrich), and primary lung fibroblasts.