GSM7595834: Adrenal Medulla P15; Mus musculus; RNA-Seq
Mouse adrenals were dissected on ice, surrounding fat and excess adrenal cortex were removed manually. Medullae were placed in an enzymatic digestion mix containing 50μg/ml DNAse I (Sigma, D5025), 10mg/ml Collagenase II (Worthington, LS004177), 2.5μg/ml Fungizone (Gibco, 15290026), 0.1X Trypsin-EDTA (Sigma, 59418C) in 1X Hank's Balanced Salt Solution (HBSS) (Gibco, 14025050) and incubated at 37°C for 15 minutes. Enzymes were inactivated by addition of 10 times volume of serum-containing Base Media: DMEM/F-12 (Gibco, 31330-038) + 5% FBS (Merk, F0804) + 50U/ml Penicillin-Streptomycin (Gibco, 15070063). The cell suspension was centrifuged at 281g for 5 minutes at room temperature, washed twice in PBS and pellets were resuspended in a solution of HBSS 2.5% FBS. An aliquot of 10,000 viable cells were used for the experiment. For Sox2 enriched dataset, cells were sorted for GFP expression. Following manifacturer's indications for 10X Genomics Single Cell RNAseq 3' v3.1