I am trying to isolate exosomes from the cell culture supernatant of Adipose Mesenchymal Stem Cells (20-30 ml) by the PEG isolation method. But exosome pellet is not visible after isolation. DLS shows the particles with sizes of 3.311nm (42%), 303nm (30%), and 5200nm (18%). Anyone here has faced the same issue. How to solve that issue. Need help in this regard.
Isolation of exosomes from cell culture media, particularly from a starting volume of around 30 ml, typically does not result in a visible pellet after isolation. This can be attributed to the small size and low density of exosomes. Methods such as polyethylene glycol (PEG) precipitation-based isolation might yield a pellet, but it can also introduce PEG contaminants, which may manifest in dynamic light scattering (DLS) results as higher size ranges. If you're encountering issues with PEG contaminants affecting your DLS results, exploring alternative isolation methods is advisable. Initially, I experimented with kit-based methods, PEG precipitation, as well as ultracentrifugation (UC). Currently, I've opted to utilize the UC method for my ongoing experiments due to its efficacy and minimal contamination concerns.
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