Isolation of mitochondria with high respiratory control from primary cultures of neurons and astrocytes using nitrogen cavitation
Isolation of mitochondria with high respiratory control from primary cultures of neurons and astrocytes using nitrogen cavitation
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DOI：
10.1016/j.jneumeth.2005.08.018
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发表时间：
2006-04-15
影响因子：
3
通讯作者：
Fiskum, G
中科院分区：
文献类型：
--
作者：
Kristián, T;Hopkins, IB;Fiskum, G
为了研究神经元或胶质细胞特异性线粒体，需要从原代神经元或星形胶质细胞培养物中分离这些细胞器。本研究为从神经元和星形胶质细胞中分离功能完整、形态完整的线粒体提供了新的方法。在第一步中，通过使用氮气空化技术破坏细胞膜从细胞中释放线粒体。该技术基于从压力容器内快速减压细胞悬浮液。然后通过Percoll梯度离心将从细胞体释放的线粒体与其余的细胞匀浆分离。这是一种相对快速的技术，其产生非常良好偶联的线粒体，其表现出与使用常规技术从脑组织分离的线粒体相当的功能和形态特征。因此，这种技术将允许检查线粒体，这是专门的细胞特异性起源。(c)2005 Elsevier B.V.保留所有权利。
To study neurons or glia-specific mitochondria one needs to isolate these organelles from primary neuronal or astrocytic cell culture. This work provides novel method for isolation of functional and morphologically intact mitochondria from neurons and astrocytes in cell cultures. In the first step, mitochondria are released from cells by disruption of cell membranes using a nitrogen cavitation technique. This technique is based on rapid decompression of a cell suspension from within a pressure vessel. Mitochondria released from cell bodies are then separated from the rest of cell homogenate by Percoll gradient centrifugation. This is a relatively rapid technique that yields to very well coupled mitochondria that exhibited functional and morphological characteristics comparable to mitochondria isolated from brain tissue using common techniques. This technique thus will allow examination of mitochondria that are exclusively cell specific in origin. (c) 2005 Elsevier B.V. All rights reserved.