GSM6254905: Human, replicate 2, scRNAseq; Homo sapiens; RNA-Seq
All human tissue specimens were collected with institutional review board (IRB) approval. The tissue was processed immediately upon isolation to ensure highest possible cell quality. Murine samples were cut into fine pieces and digested in Medium 199 (M199, Gibco) with 2% (v/v) fetal bovine serum (FBS, Gibco), Liberase (0.5WU/ml, Roche) and DNAse I (0.1 KU, ThermoFisher) 3 x 15 minutes at 37°C under constant agitation. Human samples were processed by digestion with M199 with 2% FBS, DNAse I (0.1 KU) and 2 mg/ml Stemxyme 1 (Worthington) for 2 x 30 minutes at 37°C under constant agitation. For the last 30 minutes the samples were digested with the Stemxyme/DNAse I cocktail in combination with 0.125% Trypsin (Gibco). All samples were digested in the presence of RNase inhibitors (RNasin (Promega) and RNase OUT (Invitrogen). Sorted thymus stromal cells were encapsulated into emulsion droplets using the Chromium Controller (10X Genomics). scRNA sequencing libraries were subsequently prepared using Chromium Single Cell 3 v2 and v3 Reagent kits (10X Genomics). Sorted thymus stromal cells were encapsulated into emulsion droplets using the Chromium Controller (10X Genomics). scRNA sequencing libraries were subsequently prepared using Chromium Single Cell 3 v2 and v3 Reagent kits (10X Genomics).