Isolation of Normal and Cancer-Associated Fibroblasts.

Cancer-associated fibroblasts (CAFs) play important roles in regulating tumor progression, metastasis, and response to therapies. Accurately modeling the interplay between cancer cells and the tumor microenvironment (TME) requires the use of primary cells from patient samples. Here we describe methods for the isolation of both primary CAFs and fibroblasts from omental tissue using a combination of mechanical dissociation and enzymatic digestion. Primary cells can be used for functional and mechanistic studies and may be safely cryopreserved.