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The isolation of brain extracellular vesicles (EVs), and particularly exosomes, is of the utmost importance for the understanding of their role in physiological and pathological brain processes. Moreover, the detailed characterization of biochemistry and biology of cell-specific small EVs will essentially contribute to the development of novel peripherical biomarkers that allow us to follow the progression of complex and long-lasting brain disorders, such as Alzheimer’s disease (AD). Despite growing efforts to understand the ‘mirroring relationship’ between brain pathology and brain-derived EVs collected peripherally, isolating EVs directly from brain tissue remains challenging. For instance, current methods for the isolation of small EVs (e.g., exosomes) from the brain rely on mechanic and enzymatic digestion of brain tissue, which can disrupt the plasma membrane and contaminate the EV fraction with intraluminal vesicles that would not have been released otherwise. To overcome this drawback, we have developed a novel method for the isolation of brain EVs based on their spontaneous release by the tissue. Thus, the goal of this chapter is to provide methodological guidelines and details of this novel method that produces EV yield enriched in small EVs such as exosomes highlighting the advantages and limitations of this approach.

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Isolation of Spontaneously Released Extracellular Vesicles from Human and Mouse Brain

  • 实验方案

  • 发表日期：

    2026年

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  • Patrícia A. Gomes,
  • Kalliopi Skourti,
  • Filippos Katsaitis,
  • Cristian Bodo,
  • Minghao Chen,
  • Clarissa L. Waites,
  • Christos Gatsogiannis,
  • Nuno Sousa,
  • Bruno Costa-Silva,
  • Ioannis Sotiropoulos

摘要

The isolation of brain extracellular vesicles (EVs), and particularly exosomes, is of the utmost importance for the understanding of their role in physiological and pathological brain processes. Moreover, the detailed characterization of biochemistry and biology of cell-specific small EVs will essentially contribute to the development of novel peripherical biomarkers that allow us to follow the progression of complex and long-lasting brain disorders, such as Alzheimer’s disease (AD). Despite growing efforts to understand the ‘mirroring relationship’ between brain pathology and brain-derived EVs collected peripherally, isolating EVs directly from brain tissue remains challenging. For instance, current methods for the isolation of small EVs (e.g., exosomes) from the brain rely on mechanic and enzymatic digestion of brain tissue, which can disrupt the plasma membrane and contaminate the EV fraction with intraluminal vesicles that would not have been released otherwise. To overcome this drawback, we have developed a novel method for the isolation of brain EVs based on their spontaneous release by the tissue. Thus, the goal of this chapter is to provide methodological guidelines and details of this novel method that produces EV yield enriched in small EVs such as exosomes highlighting the advantages and limitations of this approach.