通过不同的技术进行细胞外囊泡隔离和表征的方法基础
Farhang Aliakbari1, Noah B Stocek2, Maxximuss Cole-André2
1Molecular Medicine Group, Robarts Research Institute, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario N6A 3K7, Canada.
Biology methods & protocols
|March 1, 2024
概括
研究人员现在可以访问四个可适应的协议,以从细胞培养中分离细胞外囊泡 (EV) 和外丰富分离物. 这些方法,包括聚合物沉和过,为各种实验室需求提供了灵活性.
科学领域:
- 生物技术是生物技术.
- 细胞生物学 细胞生物学
- 生物化学 生物化学
背景情况:
- 细胞外囊泡 (EVs) 对于细胞间通信至关重要.
- 研究需要标准化和可访问的EV隔离方法.
- 目前的隔离技术在复杂性和有效性方面各不相同.
研究的目的:
- 介绍四个不同的,用户友好的协议,用于隔离细胞培养衍生的EVs/外体.
- 为各种实验室环境提供适应性的方法.
- 根据特定的研究条件,促进选择合适的EV隔离技术.
主要方法:
- 用聚乙烯糖醇和化沉聚合物,然后进行低速离心.
- 条件化细胞培养基的超离心.
- 使用100kDa排除过器对条件介质进行过.
- 使用商业套件进行隔离.
主要成果:
- 详细介绍了四个协议,其复杂程度各不相同.
- 这些方法使用聚合物沉,过,超离心和尺寸排除色谱 (SEC).
- 净化后的电动汽车可以在4°C短期存储,也可以在-80°C长期存储.
结论:
- 提出的协议为研究人员提供了EV和外体隔离的简单选择.
- 这些方法可以通过诸如糖密度梯度离心或SEC等技术进一步细化.
- 该研究旨在使研究人员能够选择最适合他们特定实验环境的隔离方法.
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