title: High-content high-throughput assays for characterizing the viability and morphology of human iPSC-derived neuronal cultures.
abstract: Abstract Development of quantitative high-throughput in vitro assays that enable assessment of viability and morphological changes in neuronal cells is an active area of investigation in drug discovery and environmental chemical safety assessment. High-content imaging is an emerging and efficient tool for generating multidimensional quantitative cellular readouts; in addition, human induced pluripotent stem cell (iPSC)-derived neurons are a promising in vitro model system that emulates both the functionality and behavior of mature neurons, and they are available in quantities sufficient for screening workflows. The goal of this study was to develop high-content imaging and analysis methods to assess multiple phenotypes in human iPSC-derived neuronal cells. Specifically, we optimized cell culture, staining, and imaging protocols in a 384-well assay format and improved laboratory workflow by designing a one-step procedure to reduce assay time and minimize cell disturbance. Phenotypic readouts include quantitative characterization of neurite outgrowth and branching, cell number and viability, as well as measures of adverse effects on mitochondrial integrity and membrane potential. To verify the robustness of the workflow, we tested a series of compounds that are established toxicants. We report concentration-response effects of selected test compounds on human iPSC-derived neuronal cells and illustrate how the proposed methods may be used for high-content high-throughput compound toxicity screening and safety evaluation of drugs and environmental chemicals.
authors.affiliation: 1 Molecular Devices, LLC , Sunnyvale, California.
authors.forename: Oksana
authors.initials: O
authors.lastname: Sirenko
authors.forename: Jayne
authors.initials: J
authors.lastname: Hesley
authors.forename: Ivan
authors.initials: I
authors.lastname: Rusyn
authors.forename: Evan F
authors.initials: EF
authors.lastname: Cromwell
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods; D002470:Cell Survival / Q000502:physiology; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000737:chemistry / Q000166:cytology* / Q000502:physiology*; D009474:Neurons / Q000737:chemistry / Q000166:cytology* / Q000502:physiology*