Collect conditioned media from cancer cells and add it to your fibroblasts - you will get CAFs

Thank you Arseniy. How about direct co-culture using transwell ? For how long do I need to incubate them ? Do CAFs turn back to normal fibriblast when doing single culture ?

Ahmed Alfahdawi You can do co-culture too, which will take more time to activate CAFs. At least 24 hours but may be longer. If you leave your fibroblasts alone, I suppose they will get back to normal phenotype in next couple of days. Depends on how fast they divide.