title: GSM4827180: SSc-ILD S19578 CD45+; Homo sapiens; RNA-Seq
library_construction_protocol: After bronchoalveolar lavage, fresh lung explant tissue was stored in complete media on wet ice overnight. The tissue was washed in HBSS and then thoroughly minced in digestion buffer (HBSS, 2.5mg/mL Collagenase D, 100ug/mL DNAse). Minced tissue was rocked 45 minutes at 37C. Residual tissue material was transferred into fresh digestion buffer and rocked another 45 minutes at 37C. Single cells from both rounds of digest were combined and utilized for downstream analyses. Single cell preparations were labeled with a cocktail of fluorescently labeled antibodies including, CD45(BUV421), EpCAM(PE) , CD31(PE-CY7) , CD90(APC), Live/Dead(eFluor 780), and then subjected to fluorescence-activated cell sorting (FACS) to isolate specific populations. Single-cell RNA-seq library preparation was performed using the 10x Genomics Chromium platform with the Single Cell 3' Library and Gel bead kit v2 following the manufacturer's user guide.