Pluripotent stem cell derived human neuronal progenitor cells (hPSC-NPCs) and their mature neuronal cell culture derivatives may efficiently be used for central nervous system (CNS) drug screening, including the investigation of ligand-induced calcium signalization. We have established hippocampal NPC cultures derived from human induced PSCs, which were previously generated by non-integrating Sendai virus reprogramming. Using established protocols these NPCs were differentiated into hippocampal dentate gyrus neurons. In order to study calcium signaling without the need of dye loading, we have stably expressed an advanced calcium indicator protein (GCaMP6fast) in the NPCs using the Sleeping Beauty transposon system. We observed no significant effects of the long-term GCaMP6 expression on NPC morphology, gene expression pattern or neural differentiation capacity. In order to compare the functional properties of GCaMP6-expressing neural cells and the corresponding parental cells loaded with calcium indicator dye Fluo-4, a detailed characterization of calcium signals was performed. We found that the calcium signals induced by ATP, glutamate, LPA, or proteases - were similar in these two systems. Moreover, the presence of the calcium indicator protein allowed for a sensitive, repeatable detection of changes in calcium signaling during the process of neurogenesis and neuronal maturation.

多能干细胞衍生的人神经元祖细胞（hPSC-NPC）及其成熟的神经元细胞培养衍生物可有效地用于中枢神经系统（CNS）药物筛选，包括研究配体诱导的钙信号传导。我们已经建立了人类诱导的PSC衍生的海马NPC培养物，该培养物以前是通过非整合仙台病毒重编程生成的。使用已建立的协议，这些NPC分化为海马齿状回神经元。为了研究钙信号而无需添加染料，我们使用“睡美人”在NPC中稳定表达了一种先进的钙指示剂蛋白（GCaMP6fast）。转座子系统。我们没有观察到长期GCaMP6表达对NPC形态，基因表达模式或神经分化能力的重大影响。为了比较表达GCaMP6的神经细胞和负载钙指示剂Fluo-4的相应亲代细胞的功能特性，对钙信号进行了详细的表征。我们发现，由ATP，谷氨酸，LPA或蛋白酶诱导的钙信号在这两个系统中相似。此外，钙指示剂蛋白的存在允许在神经发生和神经元成熟过程中敏感，可重复地检测钙信号的变化。