Expanded adipose-derived mesenchymal stem cells (Cx601) (TiGenix NV)

Subdermal adipose tissue is liposuctioned from healthy donors. The cells are isolated by digesting the adipose tissue with type I collagenase, followed by centrifugation. The cell obtained pellet is resuspended and lysed in erythrocyte lysis solution and centrifuged. The stromal vascular fraction, resulting from the cell pellet, is placed in cell culture containers in culture medium and antibiotics, and incubated at 37 °C and 5 % CO₂ and in a humidified atmosphere. At 24–48 h post-plating, the culture medium is removed to eliminate the non-adherent cell fraction. The cells adhere to the plastic culture plates and every 3–4 days, the culture medium is changed. After reaching 90–95 % confluence, the cells are detached with trypsin/EDTA, collected, centrifuged, and expanded, without antibiotics, to the required duplication.