来自骨髓衍生的巨细胞的小细胞外囊的类的鉴定
Jiale Cheng1, Jiong Zhu1, Yujiao Liu2
1School of Basic Medical Sciences, Anhui Medical University; State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Beijing Institute of Lifeomics.
Journal of visualized experiments : JoVE
|July 17, 2023
概括
这项研究详细介绍了一种新协议,用于分离小细胞外囊泡 (sEVs) 并使用液体染色学-双重质谱法 (LC-MS/MS) 分析它们的含量. 在巨细胞中发现了数百种sEV衍生的.
科学领域:
- 生物化学 生物化学
- 细胞生物学 细胞生物学
- 蛋白质组学是指蛋白质组学.
背景情况:
- 小细胞外囊泡 (sEVs) 是细胞间通信的关键媒介,携带各种生物分子,如蛋白质,DNA和RNA.
- sEVs在调节基因表达,细胞增殖,生存,免疫力和炎症方面发挥着至关重要的作用.
- 目前对sEV的隔离方法包括超离心,这被认为是黄金标准.
研究的目的:
- 建立一个协议,将sEV从骨髓衍生的巨细胞中分离出来.
- 为了识别和表征这些孤立的sEVs内部的内源性 (peptidome).
- 为了利用液体染色学-并联质谱学 (LC-MS/MS) 进行全面的体分析.
主要方法:
- 差异超离心法用于隔离sEVs.
- 从分离的SEV中进行提取.
- 液体染色学-双重质谱法 (LC-MS/MS) 用于的识别.
主要成果:
- 该协议成功地将sEV从骨髓衍生的巨细胞中分离出来.
- 数百种源自sEVs的被确定.
- 体分析提供了关于sEVs分子载荷的见解.
结论:
- 开发的协议是有效的sEV隔离和随后的体分析.
- 这种方法可以识别许多sEV衍生的.
- 这些发现有助于通过它们的含量来理解sEVs的功能作用.
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