治疗应用的细胞外囊泡隔离过程的比较
Soraya Williams1, Maria Fernandez-Rhodes1, Alice Law2
1School of Sport, Exercise and Health Sciences, Loughborough University, Loughborough, UK.
Journal of tissue engineering
|May 30, 2023
概括
选择正确的细胞外囊泡 (EV) 隔离方法对于治疗应用至关重要. 尺寸排除色谱 (SEC) 产生了更纯的EV,但可扩展性仍然是临床翻译的挑战.
科学领域:
- 生物技术是生物技术.
- 纳米医学是一种纳米医学.
- 细胞生物学 细胞生物学
背景情况:
- 细胞外囊泡 (EVs) 显示出治疗的前景,但临床转化受到非最佳隔离技术的阻碍.
- 目前的隔离方法在它们对EV纯度和产量的影响上有很大差异.
研究的目的:
- 评估常见的隔离方法如何影响细胞外囊泡 (EV) 的纯度和产量.
- 为了比较不同的EV隔离技术,包括超离心 (UC),聚乙烯糖醇沉,全外体隔离反应剂,水性双相系统 (ATPS) 和大小排除色谱 (SEC).
主要方法:
- 使用超离心 (UC),聚乙烯甘醇 (PEG) 沉,全外体隔离反应剂和带有和没有洗或大小排除色谱 (SEC) 的水性双相系统 (ATPS) 隔离了EV.
- 使用各种方法评估纯度和表面标记物表达 (Alix,Annexin A2,CD9,CD63,CD81),包括高分辨率纳米流细胞计.
- 一项调查评估了每种方法的可扩展性和成本等务实考虑因素.
主要成果:
- 所有方法都检测到类似EV的粒子,但纯度和表面标记物表达有显著的差异.
- 标准纯度评估 (例如,颗粒与蛋白质的比率) 与定量表面标记物分析没有一致的相关性.
- 与其他方法相比,尺寸排除色谱 (SEC) 产生了较少的颗粒,但与拉斯帕宁标志物阳性度较高.
- UC和SEC因效率而受到青,但它们的可扩展性给治疗应用带来了挑战.
结论:
- 隔离方法对电动汽车的纯度和产量有很大的影响,需要对特定应用进行仔细的选择.
- 为了准确的EV评估,先进的,特定的表征方法是必不可少的,因为标准方法可能会误导.
- 标准化和可重复的EV表征对于推进治疗研究和临床转化至关重要.
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