Generation of human ALI tumor colorectal organoids. Human tumor colon tissue fragments were seeded in collagen gels under an ALI microenvironment and cultured in ISCs media. 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 ((c), n = 4). ∗
P < 0.05 versus passage 1 day 1. Representative phase contrast images of human ALI tumor colorectal organoids from four patients (T1, T2, T3, and T4) are shown (c). Scale bar: 200 μm. H&E staining of primary tumors and the tumor organoids from four patients (T1, T2, T3, and T4) are shown (d). Representative photomicrographs were shown (n = 4). Culture days 14–21. Scale bar: 100 μm.