mRNA sequencing analysis of cultured human primary airway epithelial cells upon exposure to PMN exosomes

We analyzed the mRNA expression of monolayers of human primary airway epithelial cells after exposure to 1X10^5 exosomes from activated or quiescent PMNs along with PBS treated controls Activated PMN exosome (n=3), quiescent PMN exosome (n=3) or PBS (n=2) exposed primary human airway cells were grown and terminally differentiated to confluence under polarizing air-liquid interface conditions in a 5% CO2 incubator. Each 1.9 cm2 well containing approximately 100,000 cells was administered either 1μL of PBS, or 1X105 activated or quiescent exosomes (in 1 μL PBS) to the apical surface. After 24 hours, cells were scraped and lysed and total mRNA was harvested with RNEasy kit (Qiagen) per manufacturer’s instructions.