Sandström J, Eggermann E, Charvet I, Roux A, Toni N, Greggio C, Broyer A, Monnet-Tschudi F, Stoppini L
Department of Physiology, University of Lausanne, SCAHT, Switzerland.
Hepia, Geneva, SCAHT, Switzerland.
Toxicol In Vitro. 2017 Feb;38:124-135. doi: 10.1016/j.tiv.2016.10.001. Epub 2016 Oct 8.
Alternative models for more rapid compound safety testing are of increasing demand. With emerging techniques using human pluripotent stem cells, the possibility of generating human in vitro models has gained interest, as factors related to species differences could be potentially eliminated. When studying potential neurotoxic effects of a compound it is of crucial importance to have both neurons and glial cells. We have successfully developed a protocol for generating in vitro 3D human neural tissues, using neural progenitor cells derived from human embryonic stem cells. These 3D neural tissues can be maintained for two months and undergo progressive differentiation. We showed a gradual decreased expression of early neural lineage markers, paralleled by an increase in markers specific for mature neurons, astrocytes and oligodendrocytes. At the end of the two-month culture period the neural tissues not only displayed synapses and immature myelin sheaths around axons, but electrophysiological measurements also showed spontaneous activity. Neurotoxicity testing - comparing non-neurotoxic to known neurotoxic model compounds - showed an expected increase in the marker of astroglial reactivity after exposure to known neurotoxicants methylmercury and trimethyltin. Although further characterization and refinement of the model is required, these results indicate its potential usefulness for in vitro neurotoxicity testing.
对更快速的化合物安全性测试的替代模型的需求日益增加。随着使用人类多能干细胞的新兴技术出现，生成人类体外模型的可能性引起了关注，因为与物种差异相关的因素可能被潜在消除。在研究化合物的潜在神经毒性作用时，同时拥有神经元和神经胶质细胞至关重要。我们已经成功开发出一种方案，利用源自人类胚胎干细胞的神经祖细胞生成体外三维人类神经组织。这些三维神经组织可以维持两个月并进行渐进性分化。我们发现早期神经谱系标志物的表达逐渐降低，同时成熟神经元、星形胶质细胞和少突胶质细胞特异性标志物的表达增加。在两个月的培养期结束时，神经组织不仅显示出轴突周围的突触和未成熟的髓鞘，而且电生理测量还显示出自发活动。神经毒性测试——将非神经毒性模型化合物与已知神经毒性模型化合物进行比较——显示，在暴露于已知神经毒性物质甲基汞和三甲基锡后，星形胶质细胞反应性标志物出现了预期的增加。尽管该模型需要进一步表征和完善，但这些结果表明其在体外神经毒性测试中的潜在用途。
