GSM8421834: SC398_4; Mus musculus; RNA-Seq
Prostate tumors from castrated PSp and PCSp GEMM mice were dissected, minced, and digested into single-cell suspension using Collagenase A (Sigma, 11088793001) at 37°C for 45 min with continuously gentle shaking.  The dissociated tissues were centrifuged at 500g for 5 min, followed by further digestion with 0.25% Trypsin (Corning, 25053CI) at 37°C for 5 min. The samples were filtered using a 70uM strainer (Falcon, 352350) and centrifuged at 500g at 4°C for 5 min, followed by incubation in 1x RBC lysis buffer (Biolegend, 420302) at RT for 5 min. After washing with cold DMEM, the cells were collected and resuspended in cold PBS containing 0.04% BSA (Ambion, AM2616),  followed by measurement of cell number and viability.  Two PSp tumors and two PCSp tumors were submitted to the Single Cell Genomics Core facility at MD Anderson for library preparation and sequencing. The scRNA-seq libraries were generated using the 10X Genomics Chromium Controller Instrument and Chromium Single Cell 30 V3 Reagent Kits (10X Genomics) according to the manufacturer's recommendations.