Data from: Functional and mechanistic neurotoxicity profiling using human iPSC–derived neural 3D cultures

Neurological disorders affect millions of people worldwide and appear to be on the rise. While the reason for this increase remains unknown, environmental factors are a suspected contributor. Hence, there is an urgent need to develop more complex, biologically relevant, and predictive in vitro assays to screen larger sets of compounds with the potential for neurotoxicity. Here, we employed a human induced pluripotent stem cell (iPSC)-based 3D neural platform composed of mature cortical neurons and astrocytes as a model for this purpose. The iPSC-derived human 3D cortical neuron/astrocyte co-cultures (3D neural cultures) present spontaneous synchronized, readily detectable calcium oscillations. This advanced neural platform was optimized for high-throughput screening in 384-well plates and displays highly consistent, functional performance across different wells and plates. Characterization of oscillation profiles in 3D neural cultures was performed through multi-parametric analysis that included the calcium oscillation rate and peak width, amplitude, and waveform irregularities. Cellular and mitochondrial toxicity were assessed by high-content imaging. For assay characterization, we used a set of neuromodulators with known mechanisms of action. We then explored the neurotoxic profile of a library of 87 compounds that included pharmaceutical drugs, pesticides, flame retardants, and other chemicals. Our results demonstrated that 57% of the tested compounds exhibited effects in the assay. The compounds were then ranked according to their effective concentrations based on in vitro activity. Our results show that a human iPSC-derived 3D neural culture assay platform is a promising biologically-relevant tool to assess the neurotoxic potential of drugs and environmental toxicants.神经系统疾病影响全球数百万人群，且发病率呈上升趋势。尽管此类上升的具体原因尚不明确，但环境因素被认为是潜在诱因之一。因此，亟需开发更为复杂、具有生物学相关性且可预测性的体外实验模型，以筛选更多具有神经毒性潜力的化合物。为此，本研究采用基于人类诱导多能干细胞（iPSC）的三维神经平台作为模型，该平台由成熟皮层神经元与星形胶质细胞构成。由iPSC诱导获得的人类三维皮层神经元/星形胶质细胞共培养体系（三维神经培养物）可产生自发同步、易于检测的钙振荡。该先进神经平台已针对384孔板的高通量筛选进行优化，且在不同孔板间均表现出高度一致的功能稳定性。本研究通过多参数分析对三维神经培养物的振荡特征进行表征，分析指标涵盖钙振荡频率、峰宽、振幅以及波形不规则性。采用高内涵成像技术评估细胞与线粒体毒性。为完成实验模型的表征，本研究使用了一组作用机制明确的神经调节剂。随后，本研究对包含药物、农药、阻燃剂及其他化学物在内的87种化合物组成的库进行了神经毒性特征分析。实验结果显示，57%的受试化合物在该模型中呈现出活性效应。研究人员基于化合物的体外活性，根据其有效浓度对受试化合物进行了排序。本研究结果表明，基于人类iPSC诱导的三维神经培养实验平台，是评估药物与环境毒物神经毒性潜力的极具应用前景的生物学相关工具。