GSM7978627: Mouse lungs, post-injury day 35, scRNA-seq; Mus musculus; RNA-Seq
Lung samples were minced, digested in 5 mg/mL each of collagenase II and IV (ThermoFisher Scientific, cat# 17101015 and 17104019) in serum-free Dulbecco's Modified Eagle Medium (DMEM) (ThermoFisher Scientific, cat# 11965092) at 37 oC for 30 min with gentle agitation, neutralized in DMEM with 10% v/v fetal bovine serum (FBS) (GeminiBio, cat# 100-106), and filtered at 100 μm. The resulting suspensions were centrifuged at 500g for 5 min to isolate cell pellets, incubated in RBC lysis buffer (Millipore Sigma, cat# 11814389001) for 5 min, neutralized in DMEM with 10% FBS, centrifuged, resuspended in PBS, and filtered at 40 μm to yield single-cell suspensions. Transcriptomic samples were prepared from pooled single-cell suspensions using the Chromium Single Cell 3ʹ GEM, Library & Gel Bead Kit v3, Chromium Single Cell B Chip Kit, and Chromium i7 Multiplex Kit (10X Genomics, cat# 1000075, 1000073, and 120262). Libraries were sequenced on a HiSeq 4000 Illumina platform targeting 50,000 reads per cell.