Bone marrow-derived mesenchymal stem cells (Cerecellgram-Stroke) (Samsung Medical Center)

Bone marrow (60mL) is harvested from iliac crests of each patient. Mononuclear cells are separated using density gradient centrifugation and plated in 175 cm² flask in low-glucose DMEM supplemented with autologous serum (10%) and gentamicin (20 μg/mL). Cells are grown in a humidified 5% (vol/vol) CO₂ and 37°C incubator. Medium is exchanged on days 5 to 7 to remove non-adherent cells. MSCs are harvested and subcultured when cultures approach 70% to 80% confluence.