title: Comparative neurotoxicity screening in human iPSC-derived neural stem cells, neurons and astrocytes.
abstract: Induced pluripotent stem cells (iPSC) and their differentiated derivatives offer a unique source of human primary cells for toxicity screens. Here, we report on the comparative cytotoxicity of 80 compounds (neurotoxicants, developmental neurotoxicants, and environmental compounds) in iPSC as well as isogenic iPSC-derived neural stem cells (NSC), neurons, and astrocytes. All compounds were tested over a 24-h period at 10 and 100 μM, in duplicate, with cytotoxicity measured using the MTT assay. Of the 80 compounds tested, 50 induced significant cytotoxicity in at least one cell type; per cell type, 32, 38, 46, and 41 induced significant cytotoxicity in iPSC, NSC, neurons, and astrocytes, respectively. Four compounds (valinomycin, 3,3',5,5'-tetrabromobisphenol, deltamethrin, and triphenyl phosphate) were cytotoxic in all four cell types. Retesting these compounds at 1, 10, and 100 μM using the same exposure protocol yielded consistent results as compared with the primary screen. Using rotenone, we extended the testing to seven additional iPSC lines of both genders; no substantial difference in the extent of cytotoxicity was detected among the cell lines. Finally, the cytotoxicity assay was simplified by measuring luciferase activity using lineage-specific luciferase reporter iPSC lines which were generated from the parental iPSC line. This article is part of a Special Issue entitled SI: PSC and the brain.
authors.affiliation: XCell Science Inc., Novato, CA, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Pei
authors.affiliation: Buck Institute for Research on Aging, Novato, CA, USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Peng
authors.affiliation: National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27713, USA.
authors.forename: Mamta
authors.initials: M
authors.lastname: Behl
authors.affiliation: National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27713, USA.
authors.forename: Nisha S
authors.initials: NS
authors.lastname: Sipes
authors.affiliation: National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27713, USA.
authors.forename: Keith R
authors.initials: KR
authors.lastname: Shockley
authors.affiliation: NxCell Science, Novato, CA, USA.
authors.forename: Mahendra S
authors.initials: MS
authors.lastname: Rao
authors.affiliation: National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27713, USA.
authors.forename: Raymond R
authors.initials: RR
authors.lastname: Tice
authors.affiliation: XCell Science Inc., Novato, CA, USA; Buck Institute for Research on Aging, Novato, CA, USA. Electronic address: xzeng@xcellscience.com.
authors.forename: Xianmin
authors.initials: X
authors.lastname: Zeng
mesh_terms: D001253:Astrocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D020258:Neurotoxicity Syndromes / Q000209:etiology / Q000378:metabolism
keywords: Astrocytes; Drug screening; Lineage-specific reporter lines; Neural stem cells; Neurons; Neurotoxicity; iPSC