Fig. 4

CAFs have substantial effects on the growth and morphology of well and moderately differentiated PDOs. A Schema of 3D mono- and direct co-culture models, and representative fluorescence images of 3D direct co-culture model: low magnified view (left), high magnified 3D reconstruction view (right). PDO (green), CAF-1 (red). Scale bar, 100 µm. B Representative fluorescence images overlaid with phase contrast images of PDOs (green) cultured with or without CAF-1, and quantification of the total area of PDOs (*P < 0.05; **P < 0.01; ***P < 0.001). C Representative high magnified fluorescent images of (B) showing morphological features. Scale bars, 100 µm. B Schema of the 3D mono-, indirect and direct co-culture models. E Representative fluorescence images of Grade1 PDOs mono-cultured and co-cultured indirectly or directly with CAF-2, and quantification of the total area of PDOs with CAF-1 and CAF-2 (***P < 0.001). F Schematic diagram of the protocol to evaluate morphological features of Grade2 PDOs in the presence or absence of CAFs. G Representative fluorescence images of Grade2 PDOs (573) mono-cultured or co-cultured with CAF-1, before sorting (left) and after sorting (right). Scale bars, 100 µm (low magnification), 50 µm (high magnification)