Secretome Collection

ADMSCs in P2 were cultured to reach 80% confluency. Then they were cultured in serum-free DMEM for 18 hours prior to the secretome collection. The culture medium was collected into a 50 mL centrifuge tube and centrifuged at 1500 rpm for 10 minutes to remove cell debris. Then the centrifuged cell supernatant was transferred into centrifugal filter units with a 3 kDa molecular mass cut-off (Amicon^® Ultra-15, Millipore) and concentrated to 50-fold. The centrifuged cell supernatant was centrifuged for 45 minutes at a speed of 4000 rcf. The concentrated solution was aliquoted into 1.5 mL tubes (Eppendorf, HK) and stored in −80°C for further use. The secretome used in the study was collected from one batch experiment.