Dear Tanja!

Please You look at the article:

https://christie.openrepository.com/bitstream/handle/10541/621986/e201900425.full.pdf?sequence=1&isAllowed=y

Stromal fibroblasts induce metastatic tumor cell clusters via epithelial–mesenchymal plasticity

CAFs (exp-CAF2 cells)

Organoid culture We seeded 1 × 105 DCIS cells onto 150 μl of Matrigel coating a 24-well plate, followed by culture with the DMEM/F12 medium with GlutaMAX supplement containing 5% FBS overnight (Figs 2C and S2A). Floating dead cells and media were removed the next day and viable cells attached to the Matrigel were covered with 50 μl of Matrigel and overlaid with media containing 5% FBS. The culture medium was changed every second day and the cells were cultured for 5 d. The Matrigel was mechanically destroyed and the organoids were collected into a 1.5-ml tube prior to being jellied using iPGell (GenoStaff) and fixed in formalin solution.

FACS analysis