Adipose-mesenchymal stromal cells (MSCs) were isolated from infrapatellar Hoffa fat, sub-cutaneous hip fat and abdominal fat. First-passage cells were used for the experiments.
Fat was digested with collagenase, at 37°C for 45 minutes and centrifuged at 600g for 10 minutes. The stromal vascular fraction was transferred to a CellStack chamber, at a density of 4,000 cells/cm2, in minimum essential medium (MEM) supplemented with platelet lysate. On day 8, the cells were trypsinized and expanded at a density of 2,000 cells/cm2 until day 14.