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Quantifying Metabolic Transfer Mediated by Extracellular Vesicles Using Exo-MFA: An Integrated Empirical and Computational Platform

  • Abhinav Achreja
  • Noah Meurs
  • Deepak NagrathEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2088)

Abstract

Extracellular vesicles (EVs) are ubiquitous nanoscale particles released from many different types of cells. They have been shown to contain proteins, DNA, RNA, miRNA, and, most recently, metabolites. These particles can travel through the intercellular space and bloodstream to have regulatory effects on distant recipients. When an EV reaches a target cell, it is taken up and degraded to release its contents for utilization within the cell. In addition to regulatory effects, EVs have been shown to supplement the high metabolic demands of recipient cells in a nutrient-deprived tumor microenvironment. We developed an integrated empirical and computational platform to quantify metabolic contribution of source cell-derived EVs to recipient cells. The versatile Exo-MFA software tool utilizes 13C stable-isotope tracing data to quantify the metabolic contributions of EVs from a source cell type on a recipient cell type. This is accomplished by creating EV-depleted culture medium, producing isotope-labeled EVs from the source cells, isolating the labeled EVs from the culture supernatant, culturing the recipient cells in the presence of the labeled EVs, and measuring the resulting metabolite levels across several time points.

Key words

Extracellular vesicles Exosomes Exo-MFA Stable-isotope tracing 13-Carbon metabolic flux analysis Multicellular metabolic flux analysis 

Notes

Acknowledgments

This work was supported by grants from the National Institute of Health (R01-CA222251, R01-CA204969, and R01-CA227622) awarded to D.N.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Abhinav Achreja
    • 1
    • 2
  • Noah Meurs
    • 1
    • 2
  • Deepak Nagrath
    • 1
    • 2
    • 3
    Email author
  1. 1.Department of Biomedical EngineeringUniversity of MichiganAnn ArborUSA
  2. 2.Biointerfaces InstituteUniversity of MichiganAnn ArborUSA
  3. 3.Department of Chemical EngineeringUniversity of MichiganAnn ArborUSA

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