Almost all of the protocols that I have located utilize sequential centrifugation to eliminate impurities such as cell organelles, extracellular matrix, and other compounds.
- The mixture underwent step-wise centrifugation with the last step being ultracentrifuged at 1000,000 x g for 60 to 90 minutes
- This leaves behind a pellet of purified exosomes with all supernatant additives like PBS removed and allows for resuspension.
I am attempting to isolate plant exosomes and have incorporated low RPM's (5000) resulting in about 3700 x g and signification at 40 hz for 3 minutes. I am concerned that I am destroying the exosomes via sonication. If anyone can provide some guidance that would be great. I do not need the last step in these protocols as the resulting supernatant and exosomes will be used topically.
Thanks, Michael
We have shown our encapsulation technology can be used to purify exosomes, albeit from stem cells but it should be possible with plant cells too. https://www.frontiersin.org/journals/pharmacology/articles/10.3389/fphar.2020.00679/full
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