BACKGROUND Extracellular vesicles (EVs) are a diverse group of membrane-bound nanovesicles potentially released by every cell. With the liver's unique ensemble of cells and its fundamental physiological tasks, elucidating the role of EV-mediated hepatic cellular crosstalk and their role in different pathologies has been gaining the attention of many scientists. SCOPE OF REVIEW The present review shifts the perspective into practice: we aim to critically discuss the methods used to purify and to biochemically analyse EVs from specific liver resident cells, including hepatocytes, hepatic stellate cells, cholangiocytes, liver sinusoidal endothelial cells, Kupffer cells, liver stem cells. The review offers a reference guide to current approaches. MAJOR CONCLUSIONS Strategies for EV isolation and characterization are as varied as the research groups performing them. We present main advantages and disadvantages for the methods, highlighting common causes for concern, such as FBS handling, reporting of cell viability, EV yield and storage, differences in differential centrifugations, suboptimal method descriptions, and method transferability. We both looked at how adaptable the research between human and rodent cells in vitro is, and also assessed how well either of them translates to ex vivo settings. GENERAL SIGNIFICANCE We reviewed methodological practices for the isolation and analysis of liver-derived EVs, making a cell type specific user guide that shows where to start, what has worked so far and to what extent. We critically discussed room for improvement, placing a particular focus on working towards a potential standardization of methods.

中文翻译：

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肝源性细胞外囊泡：逐细胞概述分离和鉴定方法。

背景技术细胞外囊泡（EV）是可能由每个细胞释放的膜结合的纳米囊泡的不同种类。凭借肝脏独特的细胞群及其基本的生理任务，阐明EV介导的肝细胞串扰的作用及其在不同病理中的作用已经引起了许多科学家的关注。综述的范围本综述将观点转移到了实践中：我们旨在批判性地讨论用于纯化和生化分析特定肝驻留细胞（包括肝细胞，肝星状细胞，胆管细胞，肝窦内内皮细胞，库普弗细胞，肝干细胞。该评论为当前方法提供了参考指南。主要结论电动汽车隔离和表征的策略与研究小组执行它们的方法不同。我们介绍了该方法的主要优点和缺点，突出了引起关注的常见原因，例如FBS处理，细胞活力报告，EV产量和贮存，差异离心法的差异，方法描述欠佳以及方法可移植性。我们既研究了人类细胞与啮齿动物细胞之间体外研究的适应性，也评估了它们中的任何一种转化为离体环境的能力。一般意义我们回顾了分离和分析肝源性电动汽车的方法学实践，制作了特定于细胞类型的用户指南，显示了从何处开始，迄今已奏效的方法和程度。我们认真地讨论了改进的空间，