从细胞培养介质中分离和描述细胞外囊泡
Maia Ross1,2, Anshul S Jadli1,2, Megan Meechem1,2
1Department of Physiology and Pharmacology, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada.
Methods in molecular biology (Clifton, N.J.)
|August 6, 2024
概括
研究人员开发了一种新方法,通过大小分离和表征细胞外囊泡 (EVs). 这种技术可以更好地理解EV在细胞通信和疾病研究中的作用.
科学领域:
- 细胞生物学 细胞生物学
- 生物化学 生物化学
- 纳米技术纳米技术
背景情况:
- 细胞外囊泡 (EVs) 越来越多地被认为是它们在细胞间通信中的作用.
- 它们对疾病病理生理学的参与和诊断的潜力是显著的研究兴趣的领域.
- 目前在EV隔离和表征方面的局限性阻碍了进展.
研究的目的:
- 开发一种高效且易于使用的协议,用于从细胞培养基中分离和表征细胞外囊泡 (EV).
- 为了使电动汽车根据其尺寸类别进行分离和研究.
- 为EV分析提供可重复的方法.
主要方法:
- 电动汽车的隔离使用差分离心和商用电动汽车沉套件的组合.
- 基于尺寸的电动汽车的分离.
- 使用微流体电阻脉冲传感 (MRPS) 进行量化和尺寸分布分析.
主要成果:
- 该协议产生了高度纯净的EV样本.
- 电动汽车通过尺寸有效地分开.
- 该MRPS方法准确量化颗粒,并评估大小分布.
- 该方法适用于来自各种培养细胞的EV.
结论:
- 该协议提供了一种可复制和可访问的方法来隔离和描述尺寸定义的EV群体.
- 它解决了电动汽车研究的关键局限性,促进了对它们在健康和疾病中的作用的进一步探索.
- 该方法支持基于EV的诊断和治疗方面的进展.
相关概念视频
01:20Overview Of Cell Separation And Isolation
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.
01:32Subcellular 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...
Differential Centrifugation
Differential centrifugation is...