Comparison of neurons derived from mouse P19, rat PC12 and human SH-SY5Y cells in the assessment of chemical- and toxin-induced neurotoxicity

Abstract

BackgroundExposure to chemicals might be toxic to the developing brain. There is a need for simple and robust in vitro cellular models for evaluation of chemical-induced neurotoxicity as a complement to traditional studies on animals. In this study, neuronally differentiated mouse embryonal carcinoma P19 cells (P19 neurons) were compared with human neuroblastoma SH-SY5Y cells and rat adrenal pheochromocytoma PC12 cells for their ability to detect toxicity of methylmercury (MeHg), okadaic acid and acrylamide.MethodsRetinoic acid-treated P19 and SH-SY5Y cells and nerve growth factor-stimulated PC12 cells, allowed to differentiate for 6 days, were exposed to MeHg, okadaic acid and acrylamide for 48 h. Cell survival and neurite outgrowth were assessed with the calcein-AM assay and fluorescence detection of antibodies against the cytoskeletal neuron-specific protein βIII-tubulin, respectively. The effects of glutathione (GSH) and the potent inhibitor of GSH synthesis buthionine sulfoximine (BSO) on the MeHg induced-toxicity were assessed using the PrestoBlue™ cell viability assay and the TMRE mitochondrial membrane potential assay.ResultsDifferentiated P19 cells developed the most extensive neuronal network among the three cell models and were the most sensitive neuronal model to detect neurotoxic effects of the test compounds. MeHg produced a concentration-dependent toxicity in differentiated P19 cells and SH-SY5Y cells, with statistically significant effects at concentrations from 0.1 μM in the P19 neurons and 1 μM in the SH-SY5Y cells. MeHg induced a decrease in the cellular metabolic activity and mitochondrial membrane potential (ΔΨm) in the differentiated P19 cells and SH-SY5Y cells, that were attenuated by GSH. Okadaic acid and acrylamide also showed statistically significant toxicity in the P19 neurons, but not in the SH-SY5Y cells or the P12 cells.ConclusionsP19 neurons are more sensitive to detect cytotoxicity of MeHg, okadaic acid and acrylamide than retinoic acid-differentiated SH-SY5Y cells and nerve growth factor-treated PC12 cells. P19 neurons are at least as sensitive as differentiated SH-SY5Y cells to detect the loss of mitochondrial membrane potential produced by MeHg and the protective effects of extracellular GSH on MeHg toxicity. P19 neurons may be a useful model to study neurotoxic effects of chemicals.

Development of an iron-selective antioxidant probe with protective effects on neuronal function

“…SH-SY5Y cells derived from a human neuroblastoma are used often as a robust in vitro model for the evaluation of neurotoxicity induced by candidate drugs [ 42 , 43 ]. To evaluate possible neurotoxic effects of CT51, we incubated cultured SH-SY5Y human neuroblastoma cells with CT51 and tested cell viability as a function of CT51 concentration.…”

Groove‐patterned surfaces induce morphological changes in cells of neuronal origin

“…While longer treatment of RA results in overall increased total number of cells, it simultaneously causes the larger population of S‐Type differentiated cells (Kovalevich & Langford, ). Neurotrophic factors such as nerve growth factor and brain‐derived neurotrophic factor can stimulate neuroblastoma cells to undergo neuronal differentiation within 6 days (Popova, Karlsson, & Jacobsson, ). Insulin‐like growth factor 1 combined with surface coated by ECM protein was also observed to induce the same differentiation within 3 days (Dwane, Durack, & Kiely, ).…”

Effect of Bortezomib on Global Gene Expression in PC12-Derived Nerve Cells

“…PC12 pheochromocytoma cells (rat, ATCC, Manassas, VA, USA) were used in this study. The selected cell line is often used as a reliable model of induced peripheral neuropathy and neurotoxicity [35][36][37][38][39]. The cells respond reversibly to nerve growth factor (NGF) by induction of the neuronal phenotype when plated on collagen IV coated culture flasks.…”

“…The medium was changed every 2-3 days. PC12 cell differentiation into neurons was conducted according to the protocol described by Popova et al [39]. PC12 cells were incubated at a density of 2x10 6 cells/well in 6-well culture plates on collagen IV coated (Corning, MO, USA) in RPMI-1640 medium supplemented with 2 mM L-glutamine, 10 mM HEPES, 1 mM sodium pyruvate, 4500 mg/L glucose, and 1500 mg/L sodium bicarbonate, streptomycin (100 µg/mL), penicillin (100 U/mL), 1% HS, and 100 ng/mL NGF (Sigma Aldrich, St. Louis, MO, USA).…”