GSM6032266: SVF from WT mouse, scRNAseq; Mus musculus; RNA-Seq
gWAT was excised, minced, and placed into 10 ml of PBS(–) containing 2 mg/ ml of type 2 collagenase (Worthington, Lakewood, NJ), 1.2 U/ ml of Dispase (Gibco, Carlsbad, CA), and 10 mM CaCl2 (Roche). Incubation was done for 20 minutes at 37°C with gentle agitation. After adding 10 ml of DMEM with 10% FBS, the lysate was centrifuged at 400×g for 5 minutes and the pellet was resuspended in 5 ml of erythrocyte lysis buffer and incubated for 5 minutes at room temperature. Then the lysate was filtered through a cell strainer (70 μm) into a 50 ml tube. Next, the lysate was centrifuged at 400×g for 5 minutes and the pellet was resuspended in 5 ml of PBS and filtered through a cell strainer (40 μm) into a 50 ml tube. The SVF cells were centrifuged at 400×g and resuspended in 500 μl of PBS for further analysis. The scRNA-seq libraries was prepared using Chromium Single Cell 3ʹ Reagent Kits v3 and Chromium controller (10x Genomics).