The effects of CAFs on organoids on a Transwell platform. (A) Schematic illustration of a Transwell culture platform for mouse cells. (B) Diameters of mouse organoids on a Transwell platform with or without CAFs (n = 6; 5 organoids for each well randomly were measured). (C) Number of mouse organoids on a Transwell platform with or without CAFs (n = 6). (D) Representative images of mono-cultured, co-cultured mouse organoids. (E) Growth of mouse liver tumor organoids determined by CellTiter (n = 9) and Alamar Blue Assay (n = 6). (F) Schematic illustration of a Transwell culture platform for human cells. (G) Diameters of human organoids on a Transwell platform with or without CAFs (n = 6; 5 organoids for each well randomly were measured). (H) Number of human organoids on a Transwell platform with or without CAFs (n = 6). (I) Representative images of mono-cultured, co-cultured human organoids on a Transwell platform. (J) Growth of human organoids determined by CellTiter and Alamar Blue Assay (n = 9). (K) The number of formed mouse tumor organoids in the presence or absence of CAFs in a Transwell system (n = 4 experimental settings with 3 biological replicates for each, Mann–Whitney U tests). (L) The size of formed mouse tumor organoids in the presence or absence of CAFs in a Transwell system (n = 4 experimental settings with 3 biological replicates for each; 5 organoids for each well randomly were measured). (M) Growth of mouse liver tumor organoids determined by Alamar Blue Assay (n = 4 experimental settings with 3 biological replicates for each). (N) The number of formed human tumor organoids in the presence or absence of CAFs in a Transwell system (n = 3 experimental settings with 3 biological replicates for each, Mann–Whitney U tests). (O) The size of formed human tumor organoids in the presence or absence of CAFs in a Transwell system (n = 3 experimental settings with 3 biological replicates for each; 5 organoids for each well were measured randomly). (P) Growth of mouse liver tumor organoids determined by Alamar Blue Assay (n = 3 experimental settings with 3 biological replicates for each). (B, C, E, G, H, J, L, M, O, and P) Data are expressed as means ± SD. Mann–Whitney U tests. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001. hCAF, human cancer associated fibroblast; hOR, human organoid; mCAF, mouse cancer associated fibroblast; mOR, mouse organoid.