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，且发病率呈持续上升态势。目前该增长趋势的具体成因仍不明朗，但环境因素被推测为潜在诱因之一。因此，亟需开发更复杂、具备生物学相关性且可实现预测性的体外检测模型，以筛选更多具有神经毒性潜力的化合物。
本研究采用基于人类诱导多能干细胞（human induced pluripotent stem cell, iPSC）的三维神经平台，该平台由成熟皮层神经元与星形胶质细胞构成，用于开展相关研究。由iPSC衍生的人类三维皮层神经元/星形胶质细胞共培养体系（三维神经培养体系）可产生自发同步且易于检测的钙振荡信号。该先进神经平台已针对384孔板高通量筛选进行优化，且在不同孔板与培养批次间均展现出高度一致的功能性能。
研究通过多参数分析对三维神经培养体系的振荡特征进行表征，涵盖钙振荡频率、峰宽、振幅及波形不规则性等指标。采用高内涵成像技术评估细胞与线粒体毒性。为完成检测方法的表征工作，研究使用了一组作用机制明确的神经调节剂。随后，研究人员对包含药物、农药、阻燃剂及其他化学物质的87种化合物库的神经毒性特征展开探索。
结果显示，57%的受试化合物在该检测体系中表现出活性效应。随后根据体外活性对应的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的三维神经培养检测平台是一种极具应用前景的生物学相关工具，可用于评估药物与环境毒物的神经毒性潜力。