Hello everyone,

I started cultivating pancreatic cancer organoids obtained from patient-derived tissues. I would like to establish co-cultures with the CAFs (I have them) derived from the same patient.

My questions are:

-is it better to seed organoids and CAFs or it is better to digest the organoids into single cells and seed them with CAFs?

-normally the ratio in the tumour between cancer cells and CAFs is 20:80 for pancreatic cancer. Do you think it is possible to obtain a cell culture with this ratio or the fibroblasts wuold outnumber the cancer cells easily if I seed them with this ratio? I read the literature and many groups are doing differently about this

-I already saw that CAFs can survive in matrigel domes with the organoid media. Do you think that when I co-cultivated CAFs and organoids I should modifiy the media composition? As mentioned before, I read the literature and many groups are doing differently about this

Thank you for your help, I would like to discuss with you the different possibilities