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Studying extracellular vesicle transfer by a Cre-loxP method
Anoek Zomer1, Sander Christiaan Steenbeek1, Carrie Maynard1
1Cancer Genomics Netherlands, Hubrecht Institute-Royal Netherlands Academy of Sciences (KNAW) and University Medical Centre Utrecht, Utrecht, the Netherlands.
Nature Protocols
|December 15, 2015
Summary
This study introduces a new method using the Cre-loxP system to track extracellular vesicle (EV) transfer between cells. This technique allows researchers to visualize and study EV communication in living organisms, overcoming previous limitations.
Area of Science:
- Cell Biology
- Biotechnology
- Molecular Biology
Background:
- Extracellular vesicles (EVs) mediate intercellular communication by transferring biomolecules.
- Studying EV transfer in vivo has been challenging due to difficulties in distinguishing EV-recipient cells.
- Existing methods lack the ability to specifically track EV uptake in living systems.
Purpose of the Study:
- To develop and present a detailed protocol for studying extracellular vesicle (EV) transfer in vitro and in vivo.
- To enable the discrimination of cells that have taken up EVs from those that have not.
- To facilitate the investigation of functional EV transfer using a novel Cre-loxP-based system.
Main Methods:
- Generation of Cre-expressing cells and Cre-reporter cells.
- Utilizing the Cre-loxP system for fluorescent marking of EV-recipient cells.
- Application of the system in both in vitro cell cultures and in vivo mouse models.
Main Results:
- Successful generation of Cre(+) and reporter(+) cell lines.
- Demonstration of fluorescent labeling in cells that have taken up EVs.
- Establishment of a reliable method for studying functional EV transfer.
Conclusions:
- The Cre-loxP system provides a robust tool for tracking extracellular vesicle (EV) uptake in real-time.
- This protocol significantly advances the study of intercellular communication via EVs in complex biological environments.
- The method allows for detailed analysis of EV transfer mechanisms and functions in both experimental settings.
