Bone marrow-derived mesenchymal stem cells (Yonsei University)

Mononuclear cells were isolated using Ficoll density centrifugation and placed in a 175 cm² flask (10⁶/ml) and cultured in Dulbecco's modified Eagle's low-glucose medium containing fetal bovine serum (10%), and penicillin–streptomycin (1%) in a humidified incubator, at 37°C under 5% CO₂. After 5 days, nonadherent cells were removed by replacing the medium. Attached cells developed into colonies within 5–7 days.