Establishment of two novel organoid lines from patients with combined hepatocellular cholangiocarcinoma.

Combined hepatocellular cholangiocarcinoma (cHCC-CCA) is a unique subtype of primary liver cancer displaying both hepatocytic and cholangiocytic differentiation. The development of effective treatments for cHCC-CCA remains challenging because of its high heterogeneity and lack of a suitable model system. Using a three-dimensional culture system, we successfully established two novel cHCC-CCA organoid lines from patients undergoing surgical resection for primary liver cancer. cHCC-CCA organoid lines were authenticated by fingerprint analysis, and their morphology, growth kinetics, and anchorage-independent growth were also characterized. Hematoxylin and eosin staining and immunohistochemical analysis showed that the cHCC-CCA organoids preserved the growth pattern, differentiation grade, and phenotypic characteristics of their parental tumors. Whole-exome sequencing demonstrated that patient-derived cHCC-CCA organoid lines retained the genetic alterations identified in their original tumors. Subcutaneous tumors developed in immunodeficient mice after injection of cHCC-CCA organoids. Histologically, the xenografts recapitulated the features of the original cHCC-CCA tumors, harboring both HCC and intrahepatic cholangiocarcinoma components within the same tumor. The establishment of patient-derived cHCC-CCA organoid lines with high tumorigenicity provides a valuable resource for the mechanistic investigation and drug development of this disease.
Competing Interests: Declarations. Conflicts of interest: Nothing to report. Ethical approval: The study involving human tissues was approved by the Ethics Committee of Nanjing Medical University, and the patients provided written informed consents. All animal experiments adhered to the guidelines of the Institutional Animal Care and Use Committee of Nanjing Medical University and were approved by the Animal Experimentation Ethics Committee of Nanjing Medical University (permission number 2019-SRFA-238).
(© 2024. The Author(s) under exclusive licence to Japan Human Cell Society.)