Mesenchymal stem cells were harvested from bone marrow and conditioned with serum from the ischemic stroke patient.
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.