GSM8730159: wildtype, scRNA-seq; Mus musculus; RNA-Seq
Pancreata from mixed bone marrow chimera mice were inflated using a 30-gauge needle by injecting 3-5 mL of HBSS containing 0.5 U/mL collagenase P solution and 10 μg/mL DNase into the bile duct. The inflated pancreas was digested at 37°C for 15 min. Digested pancreata were washed three times with 10 mL of HBSS containing 2% FBS. Each individual pancreas was then suspended in 5 mL complete RPMI 1640.  Islets were visualized under a dissecting microscope and hand-picked.  Pelleted islets were suspended in 200 μL of enzyme free cell dissociation buffer.  Cells were then washed with 500 μL of HBSS, resuspended in RPMI 1640. Islet cells were stained with antibodies for CD45.1, CD45.2, CD11c, and CD19, and sorted into CD45.1 and CD45.2 T cells on a FACSAria III (BD Biosciences). Library was performed according to the manufacter's instructions (single cell 3' v2 protocol, 10x Genomics)