Mesenchymal stem cells derived from the bone marrow of amyotrophic lateral sclerosis patients.
Mononuclear cells are isolated from bone marrow aspirates of the iliac crest, with a density gradient (Histopaque, density 1.077 g/ml) and washed twice with Dulbecco's Modified Eagle's Medium containing low glucose concentrations (DMEM-LG). Cells are cultured at a density of 2 × 10^5 cells/cm2, in DMEM-LG with fetal bovine serum (10% ), for 3 days at 37 °C in an atmosphere of 5% CO2. After removing non-adherent cells, the culture medium is changed twice a week. For further passages, the cells are detached by treatment with trypsin/ethylene diamine tetraacetic acid (0.25%), for 3 min at 37 °C, seeded at a density of 4 × 10^3 cells/cm2 and expanded to up to 80–90% confluence.