Dear all，
I am going to prepare conditioned media from cancer-associated fibroblasts receiving treatment. I searched the literature and am confused about whether I should use ultra-centrifugation to concentrate supernatants? If yes, how to choose Amicon® Ultra Centrifugal Filters?
Best wishes,
Zihan
Dear Zihan,
You do not need to ultracentrifugate. A simple centrifuge is used to remove cell debris ad then you can filter-sterilize the supernatant under vaccum. Make sure your CAFs are at confluence when you give them the stimuly to make the conditioned media. When the media is yellow (24,48 or 72h depending on the total volume of media and flask used), you can remove it, centrifuge, filter it and use it (usually at 10-50% of conditioned media on other cells). Store it in 15ml falcon at -80 for maximum 6 months.
I hope this helps
Regards
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