Bone marrow-derived mesenchymal stromal cells (Royal Perth Hospital)

Bone marrow mononuclear cells were separated by density gradient centrifugation and washed cells were resuspended in DMEM supplemented with FBS (10%) and plated at a density of 160,000 cells per cm2. Cultures were maintained at 37 C in a humidified incubator containing 5% CO2 for 18–72 h, before adherent cells were washed with PBS and medium replaced for continued culturing. Cells were grown to 80–90% confluency, with medium changes every 3–4 days and then passaged using TrypLE-Select and reseeded at a density of 5,000 cells/cm2 in CellSTACK factories.