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the co-culture contained approximately 100 to 1000 stromal cells and one or more epithelial cells; the minimal epithelial to stromal cell ratio was 2:1. in each co-culture, the primary epithelial cells and stromal cells were identified by immunofluorescence microscopy after the cells were fixed with paraformaldehyde, permeabilized, and stained. stromal cells were identified based on the presence of stromal cell-specific markers (ccl5 or fap), whereas epithelial cells were identified based on the presence of the epithelial cell-specific markers (keratin-19 and vimentin). epithelial cells were then immunostained using primary antibodies specific for the epithelial markers (keratin-19 and vimentin). finally, these two cell types were counterstained with 4,6-diamidino-2-phenylindole (dapi) to identify cell nuclei.

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