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在特定培养系统中对人诱导多能干细胞衍生神经元(iCell神经元)的形态学和功能表征。

Morphological and functional characterization of human induced pluripotent stem cell-derived neurons (iCell Neurons) in defined culture systems.

作者信息

Berry Bonnie J, Akanda Nesar, Smith Alec S T, Long Christopher J, Schnepper Mark T, Guo Xiufang, Hickman James J

机构信息

NanoScience Technology Center, University of Central Florida, 12424 Research Parkway, Suite 400, Orlando, FL, 32826.

出版信息

Biotechnol Prog. 2015 Nov-Dec;31(6):1613-22. doi: 10.1002/btpr.2160. Epub 2015 Sep 11.

Abstract

Pre-clinical testing of drug candidates in animal models is expensive, time-consuming, and often fails to predict drug effects in humans. Industry and academia alike are working to build human-based in vitro test beds and advanced high throughput screening systems to improve the translation of preclinical results to human drug trials. Human neurons derived from induced pluripotent stems cells (hiPSCs) are readily available for use within these test-beds and high throughput screens, but there remains a need to robustly evaluate cellular behavior prior to their incorporation in such systems. This study reports on the characterization of one source of commercially available hiPSC-derived neurons, iCell(®) Neurons, for their long-term viability and functional performance to assess their suitability for integration within advanced in vitro platforms. The purity, morphology, survival, identity, and functional maturation of the cells utilizing different culture substrates and medium combinations were evaluated over 28 days in vitro (DIV). Patch-clamp electrophysiological data demonstrated increased capacity for repetitive firing of action potentials across all culture conditions. Significant differences in cellular maturity, morphology, and functional performance were observed in the different conditions, highlighting the importance of evaluating different surface types and growth medium compositions for application in specific in vitro protocols.

摘要

在动物模型中对候选药物进行临床前测试成本高昂、耗时费力,而且往往无法预测药物对人类的效果。制药行业和学术界都在努力构建基于人体的体外试验平台和先进的高通量筛选系统,以提高临床前研究结果向人体药物试验的转化效率。源自诱导多能干细胞(hiPSC)的人类神经元可轻易用于这些试验平台和高通量筛选,但在将其纳入此类系统之前,仍需要对细胞行为进行可靠评估。本研究报告了一种市售的hiPSC衍生神经元iCell(®) Neurons的特性,评估了其长期活力和功能表现,以确定其是否适合整合到先进的体外平台中。在体外培养28天(DIV)的过程中,对使用不同培养底物和培养基组合的细胞的纯度、形态、存活率、特性及功能成熟度进行了评估。膜片钳电生理数据表明,在所有培养条件下,动作电位的重复发放能力均有所增强。在不同条件下观察到细胞成熟度、形态和功能表现存在显著差异,这凸显了评估不同表面类型和生长培养基成分对于特定体外实验方案应用的重要性。

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