GSM7749241: CD45+ cells, Apoe/Cd36 double-null HFD, scRNA-seq; Mus musculus; RNA-Seq
Mice were sacrificed and perfused with 10 ml PBS to remove peripheral blood from the adipose tissues. Then epididymal visceral white adipose tissue (eWAT) was isolated and washed with ice-cold PBS. eWAT was minced into 2-3 mm pieces by scissors and transferred to gentleMACS C-tubes (Miltenyi Biotec) with 1.5 ml enzymatic digestion mixture (Liberase, 0.77 mg/ml; Hyaluronidase, 0.3 mg/ml; Deoxyribonuclease, 0.3 mg/ml; BSA, 1mg/ml; CaCl2, in PBS). Tissues were digested in a gentle MACSTM Octo Dissociator with Heaters (Miltenyi Biotec) at 37°C for 30 min. Suspensions were immediately filtered with a 70 mm cell strainer (Fisher Scientific) and washed once with FACS buffer (5% FBS and 0.1% sodium azide in PBS). Then, CD45+ cells were selected by the EasySep Mouse CD45 Positive Selection Kit (STEMCELL Technologies). The CD45+ cells were then suspended in the FACS buffer, counted, and immediately loaded onto a 10X Genomics Chromium instrument according to the instructions from the Chromium Next GEM Single Cell 3' Reagent Kits v3.1 (10X Genomics, Pleasanton, CA, USA). 5,000 live cells per condition were loaded onto each GEM. library construction was perfrmed according to the instruction from the Chromium Next GEM Single Cell 3' Reagent Kits v3.1