Hi - I am going to isolate some exosomes from cell culture media soon (with cells grown in exosomes depleted FBS for 24 hours).
However I would like a positive control on the isolation and was thinking about using non-exosomes depleted FBS.
My question: Does anyone know how many bovine exosomes are present in "normal" FBS per ml?
Thanks in advance,
Gary
"Normal" FBS contains vesicles of all sizes and the number of "exosomes" varies greatly with each preparation. Therefore, a range 3-8 x 10(5) per ml may be a starting point.
Thanks for your response Divaker. Do you happen to have a paper or reference for the data?
I have found some information from Thermo Fishers Q&A page on exosomes.
https://www.thermofisher.com/uk/en/home/life-science/cell-analysis/exosomes/exosomes-webinar-q-and-a.html
According to this page the concentration 0f exosomes (commonly defined as the size range between 50-150 nm or I have seen this vary between 30-200) they say:
"From 100 μl serum you can recover ~1.5-3 x 10e11 exosomes ".
Here they are referring to the amount they can isolate using one of their serum specific isolation kits and it is a little unclear if they are referring to human, bovine or other serum. Or indeed if this is the "basal" level or just the (maximum) amount they can isolate.
Thanks again.
Gary
According to this paper (Angelini et al 2016) they detected ~1x10e9 exosomes per ml of the non-depleted FBS they were using (based upon a Eisa standard curve rather than NTA).
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4916547/
If anyone else has any other references they could add to this thread it would be greatly appreciated!
Gary
Hi we did perform a pilot experiment on the FBS using NTA.
We found approximately 4x10e9 exosomes & EVs per ml of media (containing 10% serum).
Thus the FBS we used contained around 3-4x10e10 exosomes per ml (100% FBS).
Hope this helps someone in the future!
Regards,
Gary
Did anyone try different FBS from different provider?
It looks like with the VWR 97068 you don't need that long centrifugation.
Did anyone try different FBS from different provider?
It looks like with the VWR 97068 you don't need that long centrifugation.
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