GSM5378577: BAT CD45+ cells from control mice; Mus musculus; RNA-Seq
Adipose tissues were harvested, shredded with scissors and then incubated for 30 minutes with PBS containing 1.5 mg/ml collagenase A at 37°C. Digested adipose tissues were homogenized using a 1mL syringe with a 20G needle and passed through a 100µm sieve. The resulting suspensions were centrifuged (400g, 5min, 4°C) and stained with anti CD45-APC.Cy7 antibody. Cells were washed and passed again through a 100µm sieve. Unstained cells were acquired to measure autofluorescence. Dapi was added to the cell preparation before acquisition, and CD45+ Dapi- cells were purified using a BD FACS Aria III cell sorter. Reverse transcription, cDNA synthesis/amplification and library preparation were performed according to the 10x Genomics protocol (Chromium™ Single Cell 3′ Reagent Kit, v3.1 Chemistry).