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Updated: May 11, 2026
Tracking miRNA Release into Extracellular Vesicles using Flow Cytometry
Published on: October 6, 2023
Methods of analysis of dendritic cell-derived exosome-shuttle microRNA and its horizontal propagation between
Angela Montecalvo1, Adriana T Larregina, Adrian E Morelli
1Thomas E. Starzl Transplantation Institute, Department of Surgery, University of Pittsburgh Medical Center, Pittsburgh, PA, USA.
Abstract:
Exosomes are extremely small (<100 nm) membrane vesicles, generated in the endocytic compartment that are released to the extracellular milieu by living cells. Although the biological function of exosomes in vivo remains unclear, they seem to function as mechanisms of cell-to-cell communication for horizontal transfer of proteins, antigens, prions, morphogens, mRNA, and noncoding regulatory RNAs, including microRNAs (miRNAs) (also known as exosome-shuttle miRNAs). Dendritic cells (DCs), the most potent professional antigen-presenting leukocytes of the immune system, release relatively high levels of exosomes and also interact with free exosomes present in the extracellular space. Therefore, DCs constitute a good model for the analysis of exosome-shuttle miRNAs and their horizontal propagation between cells. This chapter provides basic protocols for purification of exosomes released by mouse bone marrow-derived DCs, analysis of their miRNA content, and assessment of the function of exosome-shuttle miRNAs, once they are transferred to target/acceptor DCs.
Insights
This study details methods for isolating exosomes from dendritic cells (DCs) and analyzing their microRNA (miRNA) content. It also covers assessing the function of exosome-shuttle miRNAs transferred between DCs.
Area of Science:
- Cell Biology
- Immunology
- Molecular Biology
Background:
- Exosomes are nanoscale extracellular vesicles involved in intercellular communication.
- Dendritic cells (DCs) are key immune cells that release and interact with exosomes.
- Exosomes can transfer various molecules, including microRNAs (miRNAs), between cells.
Purpose of the Study:
- To provide protocols for exosome purification from mouse bone marrow-derived DCs.
- To outline methods for analyzing miRNA content within these exosomes.
- To describe how to assess the functional impact of exosome-shuttle miRNAs on recipient DCs.
Main Methods:
- Isolation of exosomes from mouse bone marrow-derived dendritic cells.
- Analysis of microRNA profiles within purified exosomes.
- Functional assays to evaluate the transfer and impact of exosome-shuttle miRNAs on target DCs.
Main Results:
- Established protocols for exosome purification and miRNA analysis from DCs.
- Demonstrated the feasibility of assessing exosome-shuttle miRNA function.
- Highlighted DCs as a valuable model for studying exosome-mediated intercellular communication.
Conclusions:
- Dendritic cell-derived exosomes carry functional microRNAs that can be transferred to other cells.
- The provided protocols facilitate research into exosome-shuttle miRNA mechanisms.
- Understanding exosome-miRNA transfer is crucial for deciphering intercellular communication in immunity.
