Department of Neurology and Epileptology, Hertie-Institute for Clinical Brain Research, University of Tübingen, Tübingen, Germany.
Department of Cellular Neurology, Hertie-Institute for Clinical Brain Research, University of Tübingen, Tübingen, Germany.
Elife. 2019 Sep 9;8:e48417. doi: 10.7554/eLife.48417.
Most of our knowledge on human CNS circuitry and related disorders originates from model organisms. How well such data translate to the human CNS remains largely to be determined. Human brain slice cultures derived from neurosurgical resections may offer novel avenues to approach this translational gap. We now demonstrate robust preservation of the complex neuronal cytoarchitecture and electrophysiological properties of human pyramidal neurons in long-term brain slice cultures. Further experiments delineate the optimal conditions for efficient viral transduction of cultures, enabling 'high throughput' fluorescence-mediated 3D reconstruction of genetically targeted neurons at comparable quality to state-of-the-art biocytin fillings, and demonstrate feasibility of long term live cell imaging of human cells . This model system has implications toward a broad spectrum of translational studies, regarding the validation of data obtained in non-human model systems, for therapeutic screening and genetic dissection of human CNS circuitry.
我们对人类中枢神经系统回路和相关疾病的大多数认识都来自于模式生物。这些数据在多大程度上可以转化到人类中枢神经系统中，在很大程度上仍有待确定。从神经外科切除术中获得的人类脑片培养物可能为解决这种转化差距提供新的途径。我们现在证明了在长期脑片培养物中，人类锥体神经元的复杂神经元细胞结构和电生理特性得到了很好的保留。进一步的实验描述了高效病毒转导培养物的最佳条件，使“高通量”荧光介导的基因靶向神经元的 3D 重建达到与最先进的生物胞嘧啶填充相当的质量，并证明了对人类细胞进行长期活细胞成像的可行性。这种模型系统对广泛的转化研究具有意义，可用于验证非人类模型系统中获得的数据，进行治疗筛选和人类中枢神经系统回路的遗传剖析。
