Neurotoxicity is mediated by a variety of modes-of-actions leading to disturbance of neuronal function. In order to screen larger numbers of compounds for their neurotoxic potential, in vitro functional neuronal networks (NN) might be helpful tools. We established and characterized human NN (hNN) from hiPSC-derived neural progenitor cells by comparing hNN formation with two different differentiation media: in presence (CINDA) and absence (neural differentiation medium (NDM)) of maturation-supporting factors. As a NN control we included differentiating rat NN (rNN) in the study. Gene/protein expression and electrical activity from in vitro developing NN were assessed at multiple time points. Transcriptomes of 5, 14 and 28 days in vitro CINDA-grown hNN were compared to gene expression profiles of in vivo human developing brains. Molecular expression analyses as well as measures of electrical activity indicate that NN mature into neurons of different subtypes and astrocytes over time. In contrast to rNN, hNN are less electrically active within the same period of differentiation time, yet hNN grown in CINDA medium develop higher firing rates than hNN without supplements. Challenge of NN with neuronal receptor stimulators and inhibitors demonstrate presence of inhibitory, GABAergic neurons, whereas glutamatergic responses are limited. hiPSC-derived GABAergic hNN grown in CINDA medium might be a useful tool as part of an in vitro battery for assessing neurotoxicity.

神经毒性由导致神经元功能紊乱的多种作用方式介导。为了筛查大量化合物的神经毒性潜力，体外功能神经元网络（NN）可能是有用的工具。通过将hNN的形成与两种不同的分化培养基进行比较，我们从hiPSC衍生的神经祖细胞中建立并表征了人类NN（hNN）：成熟支持因子的存在（CINDA）和不存在（神经分化培养基（NDM））。作为NN控制，我们在研究中包括了区分大鼠NN（rNN）。在多个时间点评估了来自体外发育的NN的基因/蛋白质表达和电活性。体外5、14和28天的转录组将CINDA生长的hNN与体内人类发育中的大脑的基因表达谱进行了比较。分子表达分析以及电活性的测量表明，随着时间的流逝，NN成熟为不同亚型和星形胶质细胞的神经元。与rNN相比，hNN在分化时间相同的时间内电活性较低，但是在CINDA培养基中生长的hNN的发芽率要高于不含补充剂的hNN。NN对神经元受体刺激剂和抑制剂的挑战表明存在抑制性GABA能神经元，而谷氨酸能反应受到限制。在CINDA培养基中生长的源自hiPSC的GABA能hNN可能是有用的工具，可作为评估神经毒性的体外电池的一部分。