GSM6705099: Heart_old_GEX; Mus musculus; RNA-Seq
Heart and medistinal lymph nodes were collected from mice from the different age groups and all tissues were kept in HBSS/BSA 0.5% at 4°C until processing. Heart samples were digested in collagenase II 1000U/mL and dispase II 1.2U/ml for 45 minutes, at 37ºC. After digestion, both tissues were grinded through a 30um mesh and seeded in a plate for FACS staining. Each condition was stained with a different Hashtag TotalSeq-C antibodies to enable multiplexing. Gating strategy is defined in the supplemental methods. After sorting, cells were combined at different ratios for single cell RNA sequencing application. Cells were loaded in the 10X Genomics Chromium with the aim of recovering 8,000 cells. Libraries were generated with the Chromium Next GEM Single Cell 3' Reagents Kit v3 Chemistry according to manufacturer ´s instructions.