title: Characterization of three human cell line models for high-throughput neuronal cytotoxicity screening.
abstract: More than 75 000 man-made chemicals contaminate the environment; many of these have not been tested for toxicities. These chemicals demand quantitative high-throughput screening assays to assess them for causative roles in neurotoxicities, including Parkinson's disease and other neurodegenerative disorders. To facilitate high throughput screening for cytotoxicity to neurons, three human neuronal cellular models were compared: SH-SY5Y neuroblastoma cells, LUHMES conditionally-immortalized dopaminergic neurons, and Neural Stem Cells (NSC) derived from human fetal brain. These three cell lines were evaluated for rapidity and degree of differentiation, and sensitivity to 32 known or candidate neurotoxicants. First, expression of neural differentiation genes was assayed during a 7-day differentiation period. Of the three cell lines, LUHMES showed the highest gene expression of neuronal markers after differentiation. Both in the undifferentiated state and after 7 days of neuronal differentiation, LUHMES cells exhibited greater cytotoxic sensitivity to most of 32 suspected or known neurotoxicants than SH-SY5Y or NSCs. LUHMES cells were also unique in being more susceptible to several compounds in the differentiating state than in the undifferentiated state; including known neurotoxicants colchicine, methyl-mercury (II), and vincristine. Gene expression results suggest that differentiating LUHMES cells may be susceptible to apoptosis because they express low levels of anti-apoptotic genes BCL2 and BIRC5/survivin, whereas SH-SY5Y cells may be resistant to apoptosis because they express high levels of BCL2, BIRC5/survivin, and BIRC3 genes. Thus, LUHMES cells exhibited favorable characteristics for neuro-cytotoxicity screening: rapid differentiation into neurons that exhibit high level expression neuronal marker genes, and marked sensitivity of LUHMES cells to known neurotoxicants. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: National Center for Advancing Translational Sciences (NCATS), National Institutes of Health, Rockville, MD, USA.
authors.forename: Zhi-Bin
authors.initials: ZB
authors.lastname: Tong
authors.affiliation: Centers for Alternatives to Animal Testing (CAAT) at Johns Hopkins Bloomberg School of Public Health, 615 N. Wolfe Street, Baltimore, MD, USA.
authors.forename: Helena
authors.initials: H
authors.lastname: Hogberg
authors.affiliation: National Center for Advancing Translational Sciences (NCATS), National Institutes of Health, Rockville, MD, USA.
authors.forename: David
authors.initials: D
authors.lastname: Kuo
authors.affiliation: National Center for Advancing Translational Sciences (NCATS), National Institutes of Health, Rockville, MD, USA.
authors.forename: Srilatha
authors.initials: S
authors.lastname: Sakamuru
authors.affiliation: National Center for Advancing Translational Sciences (NCATS), National Institutes of Health, Rockville, MD, USA.
authors.forename: Menghang
authors.initials: M
authors.lastname: Xia
authors.affiliation: Centers for Alternatives to Animal Testing (CAAT) at Johns Hopkins Bloomberg School of Public Health, 615 N. Wolfe Street, Baltimore, MD, USA.
authors.forename: Lena
authors.initials: L
authors.lastname: Smirnova
authors.affiliation: Centers for Alternatives to Animal Testing (CAAT) at Johns Hopkins Bloomberg School of Public Health, 615 N. Wolfe Street, Baltimore, MD, USA.|University of Konstanz, POB 600, Konstanz, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Hartung
authors.affiliation: National Center for Advancing Translational Sciences (NCATS), National Institutes of Health, Rockville, MD, USA.
authors.forename: David
authors.initials: D
authors.lastname: Gerhold
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D059290:Dopaminergic Neurons / Q000187:drug effects* / Q000473:pathology; D004785:Environmental Pollutants / Q000633:toxicity*; D015870:Gene Expression / Q000187:drug effects*; D057166:High-Throughput Screening Assays; D006801:Humans; D058953:Neural Stem Cells / Q000187:drug effects* / Q000473:pathology
chemical_list: D004785:Environmental Pollutants
keywords: LUHMES; SH-SY5Y; apoptosis; differentiation; neural stem cells; neurotoxicant; neurotoxicity