Illumina NovaSeq 6000 paired end sequencing; RNA-seq of In vitro differentiation of ciliated cells in ALI-cultured human airway epithelium
RNA-seq of In vitro differentiation of ciliated cells in ALI-cultured human airway epithelium
Primary airway epithelial cells were collected by nasal brushing. Collected cells were expanded for 1-2 passages in monolayer culture using Pneumacult Ex-Plus medium. After transfer to Transwell filters cells were grown for 2 days in submersion. Cell differentiation was induced by change to Pneumacult ALI medium and introduction of ALI. Cells were grown up to 21 days in ALI setting and collected at indicated days using Tri-Reagent (Invitrogen). Total RNA was isolated using Tri-Reagent (Invitrogen). RNA isolates were DNase treated. Libraries were generated with Illumina TruSeq Stranded mRNA Library Prep Kit.