GSM5897136: emphysema lung cells [TXP24]; Homo sapiens; RNA-Seq
Human lung distal fragments were digested with a cocktail of Collagenase Type I (225 U/ml, Thermo Fisher), Dispase (15 U/ml, Thermo Fisher) and Dnase (50 U/ml, Sigma). The cell suspension was then washed with FACS buffer (2% FBS in DMEM-F12). The mixture was passed through a 70 µm cell strainer and resuspended in RBC lysis buffer, before passing through a 40 µm cell strainer. DAPI was used to exclude dead cells. Doublets and dead cells were excluded based on forward scatter, side scatter, and DAPI fluorescence. Cells were sorted into FACS buffer. FACS analysis was performed by FACSDiva. droplet-based single cell RNA-Seq: cDNA libraries were prepared for sequencing using standard protocol for the 10x Single Cell instrument, using Single cell 3' v2 chemistry kit.