Hi all,
I need your guys' advice as to how to increase the concentration of exosomes isolated from conditioned media.
I am trying to isolate exosomes from adipose-derives stem cell. I usually collect 150 ml of conditioned media (15 ml of media per T75 flask *10) all at once after 72 hours, but this has produced a concentration of exosomes (0.1 ug/ul).
Should I collect conditioned media every 24 or 48 hours multiple times to increase the volume?
Monensin (~10uM) can increase exosome yields as long as it is not toxic to the cells.
Yinan Yuan the confluence of your cell culture plates could be the problem. What is your seeding density and how fast do your cells grow?
Also, what is the isolation method?
150ml is a reasonable volume, have you checked if you are losing them during the preparation?
Hi,
Actually, the Corning™ CELLine Disposable Bioreactor system works really well. its used for high yield Ab production but it also works very well for exosome production (ref Increased exosome production from tumour cell cultures using the Integra CELLine Culture System. J Immunol Methods. 2008 Jun 1;335(1-2):98-105 Michell et al 2008)
kind regards
ketil
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