MISEV 2018 recommands Albumin, but since i use FBS for my culture, shall i look for
Bovine Serum Albumin co_isolation?
thank you
Here is what I would propose: If possible, avoid FCS-containing media when collecting cell culture supernatant for EV purification. Bovine albumin is undoubtedly a critical contaminant, but there are several others in FCS. Is there any serum-free alternative, such as supplemented media? If so, go ahead and give it a try, ensuring that cell viability and characteristics are not altered.
If adding FCS is unavoidable, you can certainly use an anti-bovine albumin antibody for immunoblotting to access purity. Just make sure to deplete endogenous bovine EVs from your FCS beforehand and keep in mind that no EV purification/separation method is entirely efficient. You will always observe that FCS components contaminate your cell culture EVs.
Try to use the most suitable methods and culture conditions for your cells and EVs. And aim to prepare your EVs as "clean" as possible.
Best of luck with your experiments!
Thank you very much for the answer Daniel Bachurski .
Best of luck to you too !
Thank you Kenneth W Witwer
; since serum free media can stress cells, I think to rather go with FBS depleted of EVs, but some say that ultracentrifugation of FBS would also deplete it from vital nutrients necessary for cell culture and recommand a commercial Exosome-Depleted FBS. did you work with it before?Thank u
Nait Eldjoudi Amina Check our article https://pubmed.ncbi.nlm.nih.gov/30577682/ in which we showed (BSA) (Sigma A9647) possible doesn't have EV contamination.
Nait Eldjoudi Amina In commercial depleted EV FBS (Life Science), they wrote on the label of the product that it has 10% of exosome (not exosome free). You can depleted exosome by the method in this paper published in Nature journal https://www.nature.com/articles/s41556-018-0040-4. They depleted EV by centrifugation .