Bone marrow-derived mesenchymal stem cells (Royan Institute)

Bone marrow (50 ml) is harvested and added to 50 ml phosphate buffer saline (PBS), then loaded onto a Lymphodex and centrifuged at 1500 g for 20 minutes. Mononuclear cells are then gently collected and counted using a nucleocounter. Mononuclear cells are washed with PBS and plated at 10⁶ cells/cm² on 150-cm² plates, in alpha-modified eagle medium supplemented with penicillin (100 IU), streptomycin (100 IU) and Hyclone bovine serum (10%). Floating cells are removed by medium replacement 7 days thereafter.