Research Paper Volume 16, Issue 5 pp 4396—4422

Figure 2. CWH22 and CLM22 organoids maintain histological and proliferative characteristics of the corresponding tumors. (A) H&E staining of the tumor organoids showing the resemblance of the organoids to the lumens and tubular structures of the primary tumor. IHC staining demonstrated consistent positive expression of CK20 and CDX2 in the organoids and corresponding tumors (scale bars, 100 μm). (B) IHC staining of CWH22 and CLM22 organoids demonstrating the consistent expression of p53, Ki67, MLH1, PMS2, MSH2, MSH6, and Villin, with the expression in CRC and CLM tissues (Supplementary Figure 1C). Scale bars, 100 μm. (C) Representative time course of CWH22 and CLM22 organoid growth for 14 days (scale bars, 500 μm) and local field amplification (scale bars, 200 μm). (D) Analysis of the proliferation rates of CWH22 and CLM22 organoids over time using 3D cell viability tests. (E) Bright-field images showing the morphology of CWH22 and CLM22 organoids at the 2nd, 5th, 10th, and 20th passages (P2, P5, P10, and P20, respectively; scale bars, 500 μm), and the expression of CK20 and Ki67 in representative differentiated organoids from CWH22 and CLM22 (scale bars, 50 μm).