Fig. 1.

Patient-derived high-risk colorectal adenoma organoids (HRCA-PDOs) were generated with Wnt3a-deficient medium system. A Bright-field images showing 7-day culture of CA organoids. Scale bar, 100 μm. B, C The patient-derived HRCA samples were digested and divided into two parts for organoid culture with the medium with (+Wnt3a) or without Wnt3a (−Wnt3a). The representative pictures of Passage 0 (Pa0) and Passage 2 (Pa2) were demonstrated (B, left panel). The superficial area of organoids in more than ten random captures of each group were calculated for analysis (B, right panel, n = 45). The PDO cultures from Pa2 were obtained for immunofluorescent staining of c-Myc. The representative pictures were displayed. The intensity of red fluorescence was quantified for each single cell and the threshold as strong positive c-Myc staining was set as 35. The ratio of strong positive c-Myc staining for both groups (n = 6). Scale bar: 200 μm in B, magnification: × 400 in C. All data were presented as mean ± SD and subjected to Student’s t-test: *** indicates P < 0.001 as −Wnt3a versus +Wnt3a