I would like to isolate extracellular vesicles derived from rat adipose-derived stem cells supernatant via density gradient centrifugation. Which gradient medium is the best for this application ? OptiPrep (60% v/w iodixanol), PEG or sucrose? Do you have any experiences or even know the dis/advantages of one compared to the other?
thanks a lot & all the best
I have not isolated exosomes from these particular cell preparations. However, I have used OptiPrep to isolate exosomes from other rodent sources. The medium is easy to use and consistent accross preparations. It also comes sterile prepared and endotoxin-free, which was important for my particular experiments. However, your selection may be influenced by your downstream assays. Depending on what you want to do with your exosomes once you have isolated them, I would make a selection based on what others have found successful for that particular use (will minor contamination with PEG/Sucrose/iodixanol affect your downstreem assays? What about ingredients in their respective dilution buffers?). Regardless of what you choose to do, you will have to verify and characterize the exosomes from the specific fractions you chose. Minor variations in your gradient dilutions could influence the types of nanoparticles found in the respective fractions. Overall, I think all three methods are valid.
Dear Mira,
kindly peruse the following articles if they would be of assistance. Best wishes.
hey @Charles D Anderson and @Benjamin Thoha Thomas, thanks a lot for your recommendations and files ! All the best with your research & have a nice day :)
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