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.