GSM8336313: miALI_scRNA; Homo sapiens; RNA-Seq
At day 82 of culture, the matrix-shaped iALI (miALI) structure was dissociated with Versene (37°C for 10 minutes) and cells were gently detached from the fibroblast-rich Collagen Chitosan matrix by pipetting to recover the apical side with epithelial cells. To ensure that only single cells were used, cells were dissociated again with Trypsin-EDTA, filtered through a 40 μm filter with PneumaCult-ALI medium complemented with 5% fetal calf serum, and centrifuged (1200 rpm for 5 minutes). The concentration of freshly dissociated cells was adjusted to 1000 cells/μL in HBSS with 0.05% BSA. The single-cell RNA sequencing (scRNA-seq) experiments were done using a 10x Chromium Controller instrument and the Chromium Single Cell 3′ Reagent Kit V3.1 (10X Genomics). Libraries were constructed according to the manufacturer's instructions using Chromium™ Single Cell 3' v3.1 Reagent Kits and a Chromium instrument. Libraries were constructed according to the manufacturer's instructions using Chromium™ Single Cell 3' v3.1 Reagent Kits and a Chromium instrument.