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通过磁性细胞分类连续分离小鼠寡细胞,微质细胞,内皮细胞,星体细胞和神经元的协议

Samah Houmam¹, Dominika Siodlak², Kevin D Pham³

¹Aging & Metabolism Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA; Department of Biochemistry & Physiology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA; Center for Geroscience and Healthy Brain Aging, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA.

|October 11, 2025

概括

这项研究引入了一种新协议,用于分离五种不同的小鼠大脑细胞类型,包括神经元和微质细胞. 这种方法为神经疾病研究提供了详细的细胞特异性分析.

科学领域:

神经科学是一个神经科学.
细胞生物学细胞生物学
生物化学生物化学

背景情况:

研究细胞类型的特异性对于理解像阿尔茨海默氏症这样的复杂神经系统疾病至关重要.
现有的分离特定脑细胞类型的方法可能限制了全面分析.

研究的目的:

介绍一个详细的协议,以隔离五种主要的小鼠大脑细胞类型:寡细胞,微质细胞,内皮细胞,星体细胞和神经元.
为了使细胞类型特定的分析用于研究神经系统疾病.

主要方法:

该协议涉及组织均质化,碎片清除,并从单个小鼠大脑中顺序分类细胞.
细胞纯度和活力通过流细胞计和定量逆转录PCR (RT-qPCR) 得到验证.

主要成果:

成功地从单个小鼠大脑中分离了寡细胞,微质细胞,内皮细胞,星体细胞和神经元.
验证了孤立细胞群的高纯度和活力.

结论:

该协议提供了一种可靠的方法,可以从单个样本中获得多个纯脑细胞种群.
孤立的细胞适用于各种下游应用,包括基因组学,转录组学和蛋白质组学,推进神经疾病研究.

关键词:

细胞隔离细胞隔离流动细胞计量是流动细胞计量的方法.基因表达的基因表达方式神经科学神经科学

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Overview Of Cell Separation And Isolation

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Cell separation was first achieved in 1964 by S. H. Seal, who separated large tumor cells from the smaller blood cells using filtration. Two years later, Pohl and Hawk performed experiments on how cells respond differently to a nonuniform electric field based on the cell type. Such observations were the inception of cell separation methods, which allow isolating a single cell type from a heterogeneous sample.

Subcellular Fractionation

Subcellular Fractionation

The homogenate obtained after cell lysis contains various membrane-bound organelles that can be further separated into pure fractions by subcellular fractionation. These isolates are used to study specific cellular components, analyze localized protein activity, and are even employed in diagnostics. Fractionation is typically achieved using centrifugation methods, the most common being density-gradient and differential centrifugation.
Differential Centrifugation
Differential centrifugation is...