三维分化 LUHMES 细胞系,研究恢复和延迟神经毒性效应。

Georgina Harris, Helena Hogberg, Thomas Hartung, Lena Smirnova
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引用次数: 0

摘要

目前的神经毒性测试和体外神经变性分子机制研究通常侧重于化合物的急性暴露。三维伦德人间脑(LUHMES)细胞可通过长期治疗或脉冲暴露结合化合物冲洗来研究延迟神经毒性效应以及恢复和神经退行性变的途径。在本单元中,我们将介绍三维 LUHMES 培养和表征。该模型的表征包括免疫细胞化学、流式细胞术和 qPCR 测量。研究化合物的延迟效应与人类暴露和神经退行性疾病的关系更为密切,这些疾病具有很强的遗传或环境因素。大多数分子终点检测方法都是针对单层细胞培养开发的,因此需要根据三维模型进行调整。在本单元中,我们将进一步介绍针对分子终点(如 ATP 水平、线粒体活力和神经元生长)的毒理学检测方法,这些方法已被调整用于三维 LUHMES 培养物。© 2017 by John Wiley & Sons, Inc.
本文章由计算机程序翻译,如有差异,请以英文原文为准。

3D Differentiation of LUHMES Cell Line to Study Recovery and Delayed Neurotoxic Effects.

3D Differentiation of LUHMES Cell Line to Study Recovery and Delayed Neurotoxic Effects.

3D Differentiation of LUHMES Cell Line to Study Recovery and Delayed Neurotoxic Effects.

3D Differentiation of LUHMES Cell Line to Study Recovery and Delayed Neurotoxic Effects.

Current neurotoxicity testing and the study of molecular mechanisms in neurodegeneration in vitro usually focuses on acute exposures to compounds. 3D Lund human mesencephalic (LUHMES) cells allow long-term treatment or pulse exposure in combination with compound washout to study delayed neurotoxic effects as well as recovery and neurodegeneration pathways. In this unit we describe 3D LUHMES culture and characterization. Characterization of the model involves immunocytochemistry, flow cytometry, and qPCR measurements. Studying the delayed effects of compounds is more relevant to human exposures and neurodegenerative diseases with a strong genetic or environmental component. Most assays for molecular endpoints have been developed for monolayer cell culture and therefore need to be adapted for 3D models. In this unit, we further describe toxicological assays for molecular endpoints such as ATP levels, mitochondrial viability, and neurite outgrowth, which have been adapted for use in 3D LUHMES cultures. © 2017 by John Wiley & Sons, Inc.

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