Version 1
: Received: 7 May 2024 / Approved: 10 May 2024 / Online: 10 May 2024 (10:24:58 CEST)
How to cite:
Noboa, J.; Leon, J.; Castro, J.; Fletes, A.; Madrigal, P.; Álvarez, I.; Navarro, R. Comparison of Methods for Isolating Exosomes from Plasma in Subjects with Normal and High-Fat Percentage and 3T3-L1 Cell Culture. Preprints2024, 2024050665. https://doi.org/10.20944/preprints202405.0665.v1
Noboa, J.; Leon, J.; Castro, J.; Fletes, A.; Madrigal, P.; Álvarez, I.; Navarro, R. Comparison of Methods for Isolating Exosomes from Plasma in Subjects with Normal and High-Fat Percentage and 3T3-L1 Cell Culture. Preprints 2024, 2024050665. https://doi.org/10.20944/preprints202405.0665.v1
Noboa, J.; Leon, J.; Castro, J.; Fletes, A.; Madrigal, P.; Álvarez, I.; Navarro, R. Comparison of Methods for Isolating Exosomes from Plasma in Subjects with Normal and High-Fat Percentage and 3T3-L1 Cell Culture. Preprints2024, 2024050665. https://doi.org/10.20944/preprints202405.0665.v1
APA Style
Noboa, J., Leon, J., Castro, J., Fletes, A., Madrigal, P., Álvarez, I., & Navarro, R. (2024). Comparison of Methods for Isolating Exosomes from Plasma in Subjects with Normal and High-Fat Percentage and 3T3-L1 Cell Culture. Preprints. https://doi.org/10.20944/preprints202405.0665.v1
Chicago/Turabian Style
Noboa, J., Iñaki Álvarez and Rosa Navarro. 2024 "Comparison of Methods for Isolating Exosomes from Plasma in Subjects with Normal and High-Fat Percentage and 3T3-L1 Cell Culture" Preprints. https://doi.org/10.20944/preprints202405.0665.v1
Abstract
The adipose tissue is responsible for fat storage and is the major producer of exosomes. Biological content transported by the exosomes provides information, such as immunometabolic alterations. This study aimed to isolate whole exosomes from plasma and supernatant of differentiated 3T3-L1 cells using three isolation techniques. Plasma from 120 individuals categorized by body adipose tissue proportion and differentiated 3T3-L1 cells were used in this study. The exosome isolation techniques were differential centrifugation (DC), size exclusion chromatography (SEC), and a commercial kit (Invitrogen), and characterized by cryo-TEM, TEM, and western blot for the tetraspanins CD9 and CD81. In the three isolation techniques, cryo-TEM and TEM analysis showed similar quality, quantity, and morphology of exosomes; at the same time, no difference was observed between individuals categorized by proportion of body fat. According to our results, the concentration of the circulating exosomes is not different between normal and high-fat percentage individuals, but rather their composition. The SEC and kit commercial proved that compile with most of the advantages of an exosome isolation technique, nerveless we emphasize the importance of selecting the appropriate methodology depending on the specific research objectives.
Keywords
exosomes; fat percentage; 3T3-L1
Subject
Biology and Life Sciences, Life Sciences
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.