GSM5608485: Sample 1; Homo sapiens; RNA-Seq
Fresh tissue was transported in cold PBS (Sigma-Aldrich) on ice. Tissue was washed three times with PBS to remove blood, then cut into tiny pieces (~1-2 mm3) and enzymatically dissociated in 1 mg/ml Type IV Collagenase in PBS for 45 min at 37 °C. The dissociation process was carried out in a shaking incubator with a speed of 50 r.p.m. Undigested masses were removed using 70-μm strainers (Corning), and red blood cells (RBC) were removed by RBC lysis buffer (Biolegend). The dissociated cells were washed twice with PBS containing 0.04% bovine serum albumin (BSA; Sigma-Aldrich). Finally, cell viability was examined by Trypan blue (Invitrogen) staining. RNA libaries were prepared with Chromium Single cell 3' Reagent v3 Kits according to the manufacture's protocol.