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使用 Seahorse XF 分析仪评估 iPSC 衍生神经元中线粒体呼吸的方案。

Protocol for evaluating mitochondrial respiration in iPSC-derived neurons by the Seahorse XF analyzer.

机构信息

Unit of Medical Genetics and Neurogenetics, Fondazione IRCCS Istituto Neurologico Carlo Besta, 20133 Milan, Italy.

Unit of Medical Genetics and Neurogenetics, Fondazione IRCCS Istituto Neurologico Carlo Besta, 20133 Milan, Italy.

出版信息

STAR Protoc. 2024 Sep 20;5(3):103127. doi: 10.1016/j.xpro.2024.103127. Epub 2024 Jul 15.

Abstract

Here, we present a protocol describing the quantification of oxygen consumption rate (OCR) and maximal respiration rate (MRR) in living induced pluripotent stem cell (iPSC)-derived neurons using the Seahorse analyzer. We guide you through the whole process: culture amplification and seeding of neural progenitor cells (NPCs), their differentiation into neurons, and normalization of the results to cell number in the analytical phase. The assessment of cellular mitochondrial function, by analyzing mitochondrial respiration, could be useful in various diseases as well as in drug screening. For complete details on the use and execution of this protocol, please refer to Aleo et al..

摘要

在此,我们提供了一份使用 Seahorse 分析仪检测活诱导多能干细胞(iPSC)衍生神经元耗氧量(OCR)和最大呼吸速率(MRR)的定量分析方案。我们将引导您完成整个过程:神经祖细胞(NPC)的培养扩增和接种、向神经元分化,以及在分析阶段将结果标准化到细胞数量。通过分析线粒体呼吸来评估细胞线粒体功能,这在各种疾病以及药物筛选中都可能很有用。有关此方案使用和执行的完整详细信息,请参考 Aleo 等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0aa1/11283151/12ea2d54593b/fx1.jpg

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