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J Vis Exp. 2018 Nov 9;(141). doi: 10.3791/58310.

Isolation of Extracellular Vesicles from Murine Bronchoalveolar Lavage Fluid Using an Ultrafiltration Centrifugation Technique.

Author information

1
Department of Medicine, Division of Pulmonary and Critical Care, Women's Guild Lung Institute, Cedars-Sinai Medical Center; Tanyalak.Parimon@cshs.org.
2
Department of Medicine, Division of Pulmonary and Critical Care, Women's Guild Lung Institute, Cedars-Sinai Medical Center.
3
Department of Medicine, Division of Pulmonary and Critical Care, Women's Guild Lung Institute, Cedars-Sinai Medical Center; Department of Biomedical Sciences, Cedars-Sinai Medical Center.
4
Department of Medicine, Smidt Heart Institute, Cedars-Sinai Medical Center.

Abstract

Extracellular vesicles (EVs) are newly discovered subcellular components that play important roles in many biological signaling functions during physiological and pathological states. The isolation of EVs continues to be a major challenge in this field, due to limitations intrinsic to each technique. The differential ultracentrifugation with density gradient centrifugation method is a commonly used approach and is considered to be the gold standard procedure for EV isolation. However, this procedure is time-consuming, labor-intensive, and generally results in low scalability, which may not be suitable for small-volume samples such as bronchoalveolar lavage fluid. We demonstrate that an ultrafiltration centrifugation isolation method is simple and time- and labor-efficient yet provides a high recovery yield and purity. We propose that this isolation method could be an alternative approach that is suitable for EV isolation, particularly for small-volume biological specimens.

PMID:
30474626
DOI:
10.3791/58310
[Indexed for MEDLINE]

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