Comparison of Methods for Isolating Exosomes from Plasma Subjects with Normal and High Fat Percentages

Adipose tissue is responsible for fat storage and is an important producer of extracellular vesicles (EVs). The biological content of exosomes, one kind of EV, provides information on aspects such as immunometabolic alterations. This study aimed to compare three plasma exosome isolation methods—usin...

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Published inLife (Basel, Switzerland) Vol. 15; no. 3; p. 410
Main Authors Noboa-Velástegui, Jacqueline, León, Juan Carlos, Castro, Jorge, Fletes, Ana, Madrigal, Perla, Álvarez, Iñaki, Navarro, Rosa
Format Journal Article
LanguageEnglish
Published Switzerland MDPI AG 06.03.2025
MDPI
Subjects
Adipocytes
Adipose tissue
Adipose tissues
Apoptosis
Body fat
Centrifugation
Centrifuges
Chromatography
Enzymes
Ethylenediaminetetraacetic acid
exosome isolation methods
Exosomes
fat percentage
Fourier transforms
Homeostasis
Methods
MicroRNA
MicroRNAs
miRNA
Plasma
plasma exosomes
Proteins
Size exclusion chromatography
Transmission electron microscopy
Type 2 diabetes
Mexico
Washington, D.C
Spain
United States > US
plasma exosomes
fat percentage
exosome isolation methods
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Summary:Adipose tissue is responsible for fat storage and is an important producer of extracellular vesicles (EVs). The biological content of exosomes, one kind of EV, provides information on aspects such as immunometabolic alterations. This study aimed to compare three plasma exosome isolation methods—using a commercial kit (CK), size exclusion chromatography (SEC), and differential centrifugation (DC)—and select the best one. Individuals categorized by normal and high body fat percentages were used. The DC and CK were proven to be the most advantageous out of the exosome isolation methods, so we suggest these methods for further protein and molecular analyses, respectively. Still, we emphasize the importance of selecting an appropriate methodology depending on the specific research objectives. At the same time, no statistical differences in exosome quality, morphology, total protein, or microRNA concentration were observed between individuals categorized by body fat percentage, so we suggest that the exosomal cargo varies in individuals with normal and high fat percentages.
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This article is a revised and expanded version of a paper published in Noboa, J.; León, J.; Castro, J.; Fletes, A.; Madrigal, P.; Álvarez, I.; Navarro, R. Comparing three methods for the isolation of exosomes from plasma in subjects with overweight and 3T3-L1 cell culture. In Proceedings of the European Congress of Immunology, Dublin, Ireland, 1–4 September 2024.
ISSN:2075-1729
2075-1729
DOI:10.3390/life15030410