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相关概念视频

Overview Of Cell Separation And Isolation01:20

Overview 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.

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相关实验视频

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Paper-based Devices for Isolation and Characterization of Extracellular Vesicles
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对血清细胞外囊泡的有效隔离和表征方法

Hatsune Mukohda1, Muhammet Mustafa Binici1, Katsuya Miyake2

  • 1Graduate School of Science and Technology, Niigata University, Niigata, Japan.

Animal science journal = Nihon chikusan Gakkaiho
|August 8, 2025
PubMed
概括

这项研究优化了从血清中分离和识别细胞外囊泡 (EV) 的方法. 过 (FL) 已被证明是获得高度净化的肉电气体的最有效方法,符合国际标准.

关键词:
这些都是肉肉.细胞外囊泡中的细胞外囊泡.隔离隔离隔离隔离隔离蛋白质标记物是蛋白质的标记物.血液中的血清是血清.

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科学领域:

  • 兽医科学 兽医科学 兽医科学
  • 生物化学 生化学
  • 细胞生物学 细胞生物学

背景情况:

  • 细胞外囊泡 (EVs) 对于细胞间的通信至关重要.
  • 缺乏用于血清EV隔离和表征的标准化方法.
  • 目前的方法并不完全符合细胞外膀研究最小信息 (MISEV) 的指导方针.

研究的目的:

  • 建立有效的方法来隔离和表征血清EVs.
  • 评估和比较四种传统的隔离技术:超离心 (UC),两种聚合物基沉 (PP) 和过 (FL).
  • 确保孤立的电动汽车符合MISEV指南.

主要方法:

  • 使用UC,PP (两种类型) 和FL从血清中进行EV隔离.
  • 通过Western Blotting (WB) 对EV标志物 (CD9,PDCD6IP,TSG101,calnexin) 的表征.
  • 纳米粒子跟踪分析 (NTA) 和传输电子显微镜 (TEM) 测量尺寸,形态和结构.
  • 对于微RNA (miR-451) 表达的定量PCR (qPCR).

主要成果:

  • 所有方法都产生了具有EV标记蛋白的样本,颗粒大小为30-150nm,具有典型的EV形态.
  • 过 (FL) 通过WB证明了最明显的EV标记蛋白表达.
  • 与FL隔离的EV表现出miR-451的最高表达,这是一个稳定的EV相关微RNA.

结论:

  • 建立了肉EV的最佳表征方法.
  • 过 (FL) 被提出为一种优越的方法,用于从血清中分离高度净化的EV.
  • 这项研究为标准化肉电动汽车研究提供了基础.