Establishment of a Novel Model for Anticancer Drug Resistance in Three-Dimensional Primary Culture of Tumor Microenvironment

Figure 1

Generation of human air liquid interface (ALI) normal colorectal organoids. Schematic experimental design of a three-dimensional (3D) culture system. Human normal colon tissue fragments were seeded in collagen gels under an ALI microenvironment and cultured in intestinal stem cells (ISCs) media (a). Pictures were taken at days 1 and 7 after seeding tissues and at days 1 and 7 after passage (b). Scale bar: 5 mm. Enlarged pictures of postpassage are shown under the original pictures at passage 1. Arrows show the grown-up organoids. The number of organoid colonies was counted (n = 4). Representative images for phase contrast microscope (c) and hematoxylin and eosin (H&E) staining (d) of human ALI normal colorectal organoids from four patients (N1, N2, N3, and N4) are shown. Culture day 30. Scale bar: 200 m (c) and 100 m (d). Expression of an epithelial cell marker, E-cadherin (e), a goblet cell marker, MUC2 (f), a fibroblast marker, vimentin (g), and a myofibroblast marker, -smooth muscle actin (SMA) (h). Representative photomicrographs were shown (n = 4). Culture day 30. Scale bar: 50 m (e–h).

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