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使用不连续聚蔗糖梯度分离纯化功能正常的血小板线粒体。

Purification of Functional Platelet Mitochondria Using a Discontinuous Percoll Gradient.

机构信息

Department of Chemistry and Biochemistry, Universite de Moncton, Moncton, NB, Canada.

出版信息

Methods Mol Biol. 2021;2276:57-66. doi: 10.1007/978-1-0716-1266-8_4.

Abstract

The isolation of mitochondria is gaining importance in experimental and clinical laboratory settings. Of interest, mitochondria and mitochondrial components (i.e., circular mitochondrial DNA, N-formylated peptides, cardiolipin) have been involved in several human inflammatory pathologies, such as cancer, Alzheimer's disease, Parkinson's disease, and rheumatoid arthritis. While several mitochondrial isolation methods have been previously published, these techniques are aimed at yielding mitochondria from cell types other than platelets. In addition, little information is known on the number of platelet-derived microvesicles that can contaminate the mitochondrial preparation or even the overall quality as well as functional and structural integrity of mitochondria. Here we describe a purification method, using a discontinuous Percoll gradient, yielding mitochondria of high purity and integrity from human platelets.

摘要

在实验和临床实验室环境中,线粒体的分离正变得越来越重要。有趣的是,线粒体和线粒体成分(即圆形线粒体 DNA、N-甲酰化肽、心磷脂)已涉及多种人类炎症性病理,如癌症、阿尔茨海默病、帕金森病和类风湿关节炎。虽然以前已经发表了几种线粒体分离方法,但这些技术旨在从除血小板以外的细胞类型中获得线粒体。此外,关于可能污染线粒体制备物的血小板衍生微泡的数量,甚至关于线粒体的整体质量以及功能和结构完整性的信息知之甚少。在这里,我们描述了一种使用不连续 Percoll 梯度的纯化方法,从人血小板中获得高纯度和完整性的线粒体。

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