We conducted morphological assessments using bright-field and immunofluorescence staining, utilizing biomarkers, including epithelial cell markers (EPCAM), colon cancer markers (cytokeratin 20, CK20), proliferation markers (KI67), and fibroblast markers (alpha-smooth muscle actin, SMA; vimentin, VIM) (Figure 1A–C) to validate the preservation of characteristics in organoids and CAFs. This evidence concerns the gene KRT20 and malignant colon neoplasm.