Study: Multiple sclerosis blood antibodies found to be toxic to neurons CUAnschutz
The 96-well tissue culture plates and 12 mm glass coverslips in the 24-well plate were used for all cell cultures.The SH-SY5Y neuroblastoma cells were obtained from ATCC . Cells were cultured in DMEM with 10% FBS and 1X Penicillin-Streptomycin on 0.1% gelatin-coated plates, passaged with 0.05% Trypsin and 0.02% EDTA solution. Cells at passage number 3–10 were used for cytotoxicity assays.
Viability/Cytotoxicity Kit was used for quantifying cell viability . Calcein AM was used for labeling live cells and the Ethidium Homodimer-1 for labeling dead cells. The live and dead cells were quantified by green fluorescent and red fluorescent , respectively.The RealTime-Glo Annexin V Apoptosis and Necrosis Assay kit was used for evaluating apoptotic and necrotic cell death. The five components were diluted to 1x in the culture medium and added to the treatment mixtures.
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