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BrainPhys神经元培养基支持小鼠原代神经元培养中线粒体的生理功能。

BrainPhys Neuronal Media Support Physiological Function of Mitochondria in Mouse Primary Neuronal Cultures.

作者信息

Faria-Pereira Andreia, Temido-Ferreira Mariana, Morais Vanessa A

机构信息

Faculdade de Medicina, Instituto de Medicina Molecular João Lobo Antunes, Universidade de Lisboa, Lisbon, Portugal.

出版信息

Front Mol Neurosci. 2022 Jun 14;15:837448. doi: 10.3389/fnmol.2022.837448. eCollection 2022.

Abstract

neuronal cultures are extensively used in the field of neurosciences as they represent an accessible experimental tool for neuronal genetic manipulation, time-lapse imaging, and drug screening. Optimizing the cultivation of rodent primary neuronal cultures led to the development of defined media that support the growth and maintenance of different neuronal types. Recently, a new neuronal medium, BrainPhys (BP), was formulated envisioning the mimicry of brain physiological conditions and suitability for cultured human iPSC-derived neurons and rat primary neurons. However, its advantages in mouse primary neuronal cultures and its effects in neuronal bioenergetics are yet to be demonstrated. In this study, we validated the beneficial use of BP in mouse primary neuronal cultures based on the observation that neuronal cultures in BP media showed enhanced ATP levels, which increased throughout neuronal maturation, a finding that correlates with higher mitochondrial activity and ATP production at later maturation stages, as well as an increased glycolysis response on mitochondrial inhibition and increased mitochondrial fuel flexibility. Taken together, our data demonstrate that BP medium promotes mitochondrial activity along with neuronal maturation of cultures.

摘要

神经元培养物在神经科学领域被广泛应用,因为它们是用于神经元基因操作、延时成像和药物筛选的一种易于使用的实验工具。优化啮齿动物原代神经元培养物的培养方法,促使了能够支持不同神经元类型生长和维持的限定培养基的开发。最近,一种新的神经元培养基BrainPhys(BP)被配制出来,旨在模拟大脑生理条件,并适用于培养的人诱导多能干细胞衍生神经元和大鼠原代神经元。然而,它在小鼠原代神经元培养中的优势及其对神经元生物能量学的影响尚未得到证实。在本研究中,我们基于以下观察结果验证了BP在小鼠原代神经元培养中的有益作用:在BP培养基中的神经元培养物显示出ATP水平升高,且在整个神经元成熟过程中持续增加,这一发现与后期成熟阶段更高的线粒体活性和ATP产生相关,同时在受到线粒体抑制时糖酵解反应增加以及线粒体燃料灵活性增强。综上所述,我们的数据表明BP培养基可促进线粒体活性以及培养物的神经元成熟。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/edd6/9239074/5484a0b3b11f/fnmol-15-837448-g001.jpg

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