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Human co-culture model of neurons and astrocytes to test acute cytotoxicity of neurotoxic compounds

Abstract: 
Alternative methods and their use in planning and conducting toxicology experiments has become essential for modern toxicologists, thus reducing or replacing living animals. Although in vitro human coculture models allow the establishment of biologically relevant cell-cell interactions that recapitulate the tissue microenvironment and better mimic its physiology, the number of publications is limited specifically addressing this scientific area and utilizing this test method which could provide an additional valuable model in toxicological studies. In the present study, an in vitro model based on CNS cell co-cultures was implemented using a trans-well system combining human neuronal cells (SH-SY5Y cell-line) and glial cells, namely astrocytes (D384 cell-line), to investigate neuroprotection of D384 on SH-SY5Y and vice-versa. The model was applied to test acute (24-48h) cytotoxicity of three different neurotoxicants: (i) methylmercury (1-2.5 μM); (ii) Fe3O4-nanoparticles (1-100 μg/ml); (iii) methylglyoxal (0.5-1 mM). Data were compared to monocultures evaluating the mitochondrial function and cell morphology. The results clearly showed that all compounds tested affected the mitochondrial activity and cell morphology in both mono- and co-culture conditions. However, astrocytes, when cultured together with neurons, diminish the neurotoxicant-induced cytotoxic effects that occurred in neurons cultured alone, and astrocytes become more resistant in the presence of neurons. This human CNS co-culture system seems a suitable cell model to feed high-throughput acute screening platforms and to evaluate both human neuronal and astrocytic toxicity and neuroprotective effects of new and emerging materials (e.g., nanomaterials) and new products with improved sensitivity due to the functional neuron-astrocyte metabolic interactions. Key Words: SH-SY5Y neurons, D384 astrocytes, methylglyoxal, methylmercury; magnetite nanoparticles; mitochondrial function.