title: GSM8208556: Adipose_CD31negative_CD45negative_Tom_negative; Mus musculus; RNA-Seq
library_construction_protocol: Gonadal adipose tissue samples were subjected to direct mincing using a sharp scissor, followed by digestion in DMEM supplemented with 0.3 unit/ml Liberase TM (Roche) and 10% BSA at 37°C for 50 min with gentle shaking. After incubation, the resulting cell suspension was centrifuged at 400g for 3 min, and the supernatant was removed. The stromal vascular fractions (SVFs) present in the cell pellet were resuspended in DMEM and passed through 100 µm cell strainers. Subsequently, the red blood cells were lysed using RBC buffer for 5 min. The remaining cells were washed with PBS and resuspended in PBS containing 4% FBS and mouse FcR blocking reagent (MACS) at 4°C for 10 min, and further stained with anti-CD31antibodies and anti-CD45 antibody at 4°C for 30 min, washed and  passed through 70 µm cell strainers and then subjected to FACS SORP Aria (BD Biosciences) to sort out cells.The single-cell suspension of CD31- CD45- of Tom+ cells and Tom- cells were conducted to the sc-RNA seq sequencing. For the scRNA-seq library construction, we followed the manufacturer's instructions of 10x Genomics Chromium Single Cell 3′ Reagent Kit v3.1.