Abstract
Objectives
Most attention has been focused on physiologically generated membrane blebs on the cellular cortex, whereas artificial membrane blebs induced by chemicals are studied to a lesser extent.
Results
We found that exposure of HeLa human cervical cancer cells to paraformaldehyde (PFA), followed by incubation in phosphate-buffered saline (PBS) efficiently induced large membrane blebs on the cellular cortex. Intriguingly, sequential exposure of the PFA-treated cells to PBS containing dimethyl sulfoxide (DMSO) facilitated shedding of the blebs from the cellular cortex, yielding a high quantity of large extracellular vesicles in the supernatant, which was applicable to assess the potentials of compounds and proteins as membrane influencers. Similar effects of PFA and DMSO were detected on the cellular cortex of other human, mouse, and fish cells.
Conclusions
Our procedure to facilitate membrane blebbing and vesicle shedding by chemicals may be practical for the manipulation of membrane dynamics and the development of vesicle-inspired technologies using a wide variety of cell types.
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Abbreviations
- Con A:
-
Concanavalin A
- DMSO:
-
Dimethyl sulfoxide
- PFA:
-
Paraformaldehyde
- PBS:
-
Phosphate-buffered saline
- ThT:
-
Thioflavin T
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Acknowledgements
We thank all the members of the Saitoh Laboratory for helpful discussion. This work was supported by Japan Society for the Promotion of Science (JSPS) KAKENHI Grant 19J20051 (S. O.), 17H01878 (H. S.) and The Terumo Life Science Foundation (Japan) for the Promotion of Science and Technologies (H. S.). S. O. is supported by the fellowship for doctor course student from the JSPS.
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Okada, S., Fukai, Y., Yoshimoto, F. et al. Chemical manipulations to facilitate membrane blebbing and vesicle shedding on the cellular cortex. Biotechnol Lett 42, 1137–1145 (2020). https://doi.org/10.1007/s10529-020-02848-7
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DOI: https://doi.org/10.1007/s10529-020-02848-7