机械刺激,如剪切应力和皮埃佐1刺激,会产生红细胞细胞外囊泡
Gurneet S Sangha1, Callie M Weber1, Ryan M Sapp1
1Fischell Department of Bioengineering, University of Maryland, College Park, MD, United States.
Frontiers in physiology
|September 18, 2023
概括
产生纯红细胞细胞外囊泡 (RBC-EVs) 对研究至关重要. 这项研究使用皮埃佐1刺激优化了RBC-EV的产生,产生了高纯度囊泡,用于准确的机械研究.
科学领域:
- 生物技术是生物技术.
- 细胞生物学 细胞生物学
- 细胞外囊泡研究研究
背景情况:
- 为了研究,产生生理上相关的红细胞细胞外囊泡 (RBC-EVs) 是很困难的.
- 标准方法往往导致低产量或不纯的囊泡制剂.
研究的目的:
- 在体外研究产生和分离高度红细胞-EVs的方法.
- 为了优化RBC-EV生产的参数,使用剪切应力和Piezo1离子通道刺激.
- 评估RBC-EV纯度对功能结果的影响.
主要方法:
- 使用剪切应力或 piezo1 激动剂 yoda1.1. 产生红细胞-EV.
- 优化红细胞-EV生成参数,包括血红素,治疗时间和剂量.
- 隔离方法的比较:基于膜的亲和力,超过和超离心 (有/没有SEC).
- 使用压力肌图进行功能评估,以评估孤立的红细胞-EVs的血管扩张效应.
主要成果:
- 最佳的红血细胞-EV生成涉及治疗红血细胞在6%的血红素与10μM的yoda130分钟.
- 超离心证明最有效的隔离RBC-EVs,最大限度地提高产量和纯度,同时最大限度地减少血液溶解.
- 纯粹的皮埃佐1-RBC-EVs没有诱导血管扩张,与含有同隔离污染物的不纯样本不同.
结论:
- 红细胞-EV可以通过皮埃佐1刺激有效地产生,在生理流动下潜在地发生在体内.
- 描述EV生成和隔离参数对于准确的机械学研究至关重要.
- 红细胞-EV纯度显著影响生物测试中的功能结果.
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