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 have become essential for modern toxicologists, thus reducing or replacing living animals. Although in vitro human co-culture 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 central nervous system (CNS) cell co-cultures was implemented using a transwell 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 (24-48 hours) cytotoxicity of 3 different neurotoxicants: (1) methyl mercury (1-2.5 μM), (2) FeO nanoparticles (1-100 μg/mL), and (3) methylglyoxal (0.5-1 mM). Data were compared to mono-cultures 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-culture 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 (eg, nanomaterials) and new products with improved sensitivity due to the functional neuron-astrocyte metabolic interactions.

Suitability of 3D human brain spheroid models to distinguish toxic effects of gold and poly-lactic acid nanoparticles to assess biocompatibility for brain drug delivery

“…The 3D LUHMES model is a monoculture model, consisting only of dopaminergic neurons, and may display higher susceptibility to harmful agents that induce mitochondrial damage, and cytotoxicity [93]. In fact, NP cytotoxicity has shown to be lower when neurons and astrocytes are co-cultured, compared to monocultures [94]. The absence of neuroprotective glial cells, together with the known susceptibility of dopaminergic neurons to mitochondria depletion, could contribute to the toxicity observed in this model from all studied NP.…”

Ethanol Induction of Innate Immune Signals Across BV2 Microglia and SH-SY5Y Neuroblastoma Involves Induction of IL-4 and IL-13

“…After the differentiation procedure described above, media was refreshed to 10% FBS-containing DMEM and Transwell inserts (Corning CLS3452, Corning, NY, USA) were placed on top of the SH-SY5Y-containing wells and allowed to equilibrate for 30 minutes. Transwells are a well-characterized 3D model of co-culture and have been used extensively for neuronal co-cultures [40,46,47,48], including with SH-SY5Y and BV2 in particular [49]. BV2 were plated (1.5 × 10 5 cells per well) on top of the Transwell inserts; see Figure 1 for a schematic.…”

“…In this study, we examine how co-culture of BV2 microglia and SH-SY5Y neuroblastoma influences ethanol-induced innate immune signaling of these cell lines. Co-culture modules using glial cells and SH-SY5Y have been used previously in multiple disease settings [39,40]. Since SH-SY5Y adopt a dopaminergic phenotype [41] and IL-4 and IL-13 modulate dopaminergic signaling, SH-SY5Y could provide insight into responses of dopaminergic neurons.…”

Concepts toward directing human astroplasticity to promote neuroregeneration

“…Toxicity is a major barrier in the field of nanomedicine that needs to be thoroughly tested for each new nanoparticle. hPSC‐derived neural cells (Repar et al, ) or astrocyte‐neuron co‐cultures (De Simone et al, ) have been utilized as a screening platform to assess neurotoxicity. One alternative to reduce toxicity, perhaps, is the utilization of exosomes from neural cells as a source for nanoparticles, which can potentially target specific cells through incorporation of surface antibodies (Kawikova and Askenase, ).…”