Validating polycarbonate ultracentrifuge tube reuse in extracellular vesicle research

Extracellular vesicles (EVs) are tiny membrane-bound particles released by cells that play crucial roles in intercellular communication and various physiological processes. However, the isolation of EVs often relies on single-use laboratory plastics, contributing to both environmental pollution and consumable costs. In a bid to address these challenges, researchers at Harvard Medical School have developed and validated a method for cleaning and reusing polycarbonate ultracentrifuge (UC) tubes in EV proteomics experiments, aiming to promote sustainability in scientific research.

The protocol for cleaning polycarbonate UC tubes involves several key steps to ensure effective removal of contaminants and residual proteins. First, the tubes are immediately submerged in water to prevent protein drying. They are then washed with a mild detergent solution, rinsed with hot tap water, followed by demineralized water, and finally, sterilized with 70% ethanol. This meticulous cleaning process aims to maintain the integrity of the tubes while eliminating any potential sources of contamination.

To validate the effectiveness of the UC tube re-use protocol in EV proteomics experiments, the researchers conducted a series of experiments involving the isolation of EVs from cardiovascular tissue. After cleaning the used tubes, the experimental process was repeated, and the obtained pseudo-EV pellets were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS).

The results of the validation study were promising, with a significant reduction in the number of identified proteins in the pseudo-pellet compared to the previous EV isolation sample from the same tube. Moreover, comparing a cleaned tube against a blank tube revealed a high degree of similarity in the protein profiles, indicating minimal contamination from previous experiments. Importantly, chromatogram analysis confirmed the absence of polymer peaks, further validating the cleanliness of the tubes.

The development and validation of a protocol for cleaning and reusing polycarbonate UC tubes represent a significant step towards sustainability in EV research. By reducing the reliance on single-use plastics and minimizing experimental costs, this protocol offers a more environmentally friendly and cost-effective approach to EV isolation and proteomics analysis. Moving forward, the adoption of such sustainable practices in scientific research has the potential to mitigate the environmental impact of laboratory activities while advancing our understanding of EV biology.

Cahalane RME, Turner ME, Clift CL, Blaser MC, Bogut G, Levy S, Kasai T, Driedonks TAP, Nolte-‘t Hoen ENM, Aikawa M, Singh SA, Aikawa E. (2024) Polycarbonate Ultracentrifuge Tube Re-use in Proteomic Analyses of Extracellular Vesicles. J Vis Exp Mar 8;(205). [abstract]

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