Adipose-derived stromal cells were isolated from lipoaspirates.
Adipose-derived stromal cells were isolated according to the following procedures: the obtained adipose tissue was washed 3-4 times, using an equal volume of KRB solution in order to remove blood from the adipose tissue. Adding an equal volume of collagenase solution to the adipose tissue, the reaction was carried out in a water bath at 37° C. The reaction product was placed in a centrifuge tube and centrifuged at 20° C, at 1,200 rpm for 10 minutes. After removing the fat layer supernatant, the remaining collagenase solution was gently separated without being shaken. A stromal medium was added to the collagenase solution to prepare a suspension, followed by centrifugation at 20° C and 1,200 rpm, for 5 minutes. Here, the sediment was a stromal vascular fraction (SVF), while the supernatant was discarded. SVF was suspended in the stromal medium, inoculated in a culture vessel and cultured for 24 hours in an incubator at 37° C, under 5% CO2. After removing the culture medium and washing the SVF with PBS, the SVF was proliferated in a stromal medium. Stromal medium contained bFGF (1ng/ml) or EGF (5 ng/ml).