scRNA-seq of Homo sapiens: adult pancreas
All remaining tissues are collected in MACS Tissue Storage Solution (Miltenyi Biotec., Auburn, CA, USA) on ice, and processed to the single cell dissociation immediately. Specimens can be maintained in MACS Tissue Storage Solution (Miltenyi Biotec.) for 48 hours at 4C without showing background effects like cell activation or apoptosis induction. Biopsy tissues are placed on a petri dish with a small volume of enzyme mixture solution (Tumor Dissociation Kit, Miltenyi Biotec.), and minced into small pieces under 1-2 mm with surgical blades. Minced pieces are transferred to gentleMACSTM C-tube containing enzyme mixture solution. By choosing an appropriate program of gentleMACSTM Dissociator, Tissue pieces are further dissociated three times with two interval incubations for 30 minutes at 37C under continuous rotation using the MACSmix Tube Rotator. Resuspended samples are applied to a cell strainer (70 m) placed on a 15 ml cornical tube, and washed with 10 ml of RPMI1640 media. After a centrifugation at 400xg for 5 minutes, cells were resuspended in an appropriate volume of RPMI1640 media, and counted after trypan blue staining or acridine orange/ propidium iodide staining. Optionally, erythrocytes are removed by red blood cell lysis solution for 10 min before counting.