Niedringhaus Mark, Dumitru Raluca, Mabb Angela M, Wang Yuli, Philpot Benjamin D, Allbritton Nancy L, Taylor Anne Marion
1] UNC/NCSU Joint Department of Biomedical Engineering [2] UNC Neuroscience Center.
1] UNC Neuroscience Center [2] UNC Human Pluripotent Stem Cell Core [3] UNC Department of Genetics.
Sci Rep. 2015 Feb 10;5:8353. doi: 10.1038/srep08353.
The effort and cost of obtaining neurons for large-scale screens has limited drug discovery in neuroscience. To overcome these obstacles, we fabricated arrays of releasable polystyrene micro-rafts to generate thousands of uniform, mobile neuron mini-cultures. These mini-cultures sustain synaptically-active neurons which can be easily transferred, thus increasing screening throughput by >30-fold. Compared to conventional methods, micro-raft cultures exhibited significantly improved neuronal viability and sample-to-sample consistency. We validated the screening utility of these mini-cultures for both mouse neurons and human induced pluripotent stem cell-derived neurons by successfully detecting disease-related defects in synaptic transmission and identifying candidate small molecule therapeutics. This affordable high-throughput approach has the potential to transform drug discovery in neuroscience.
获取用于大规模筛选的神经元所需的精力和成本限制了神经科学领域的药物发现。为克服这些障碍，我们制造了可释放的聚苯乙烯微筏阵列，以生成数千个均匀、可移动的神经元微型培养物。这些微型培养物维持着具有突触活性的神经元，这些神经元易于转移，从而使筛选通量提高了30倍以上。与传统方法相比，微筏培养物的神经元活力和样本间一致性显著提高。我们通过成功检测突触传递中的疾病相关缺陷并鉴定候选小分子疗法，验证了这些微型培养物对小鼠神经元和人诱导多能干细胞衍生神经元的筛选效用。这种经济实惠的高通量方法有潜力改变神经科学领域的药物发现。
