MSCs were isolated from adipose tissue of the thigh of a healthy donor, after a signed consent, according to the French regulations. MSCs were obtained after the digestion of adipose tissue with collagenase NB6 (Serva electrophoresis, Heidelberg, Germany). The adipose tissue was placed in 40 mL of collagenase NB6 diluted in α-MEM medium at a final concentration of 5μg/mL in a 50 mL conical tube (Falcon, Dutscher, Bernolsheim, France). After a 2 h incubation at 37C, the digested tissue was filtered through a cell strain with pores of 100μm to remove the remaining tissue. The strained cells were then centrifuged and plated in a cell factory (Thermofisher, Nunc, Waltham, United States) for expanding in MSC culture medium composed of α-MEM (Gibco, Thermofisher, Waltham, United States) supplemented with 10% fetal bovine serum (FBS, Biowest, Nuaillé, France) and 1% antibiotic/antimycotic mix (Anti-Anti 100×, Gibco, Thermofisher, Waltham, United States).

At the second passage, cells were characterized by flow cytometry using a panel of MSC positive markers including CD29-PE (MACS Miltenyi, Bergish Gladbach, Germany), CD73-PE (cat number: 550257, bd, New Jersey, United States), CD90-FITC (cat number: 555595, bd), CD105-PE (cat number: 560839, bd), and negative markers including CD31-FITC (cat number: 555445, bd) and CD34-APC (cat number: 555824, bd). Unstained cells and cells stained with control isotypes have been used as negative control.

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