OPTIMIZATIONS OF PATIENT-DERIVED CHOLANGIOCARCINOMA EXPLANT CULTURE FOR 3D ORGANOIDS FORMATIONS

Abstract

The three-dimensional (3D) organoid technology has become more popular among researchers. Organoids are the small structure of tissues and organs that mimic the 3D architectures and functions of a specific organ. These 3D constructs represent near-physiological in vitro models and support many biomedical applications, including cancer research. Cholangiocarcinoma (CCA) is a malignant tumor of bile duct epithelium which highly endemic to the northeastern part of Thailand. CCA cell lines were established successfully as 2D cultures; however, some aspects require drug evaluation on multi-phenotypic cell types. Currently, the development of precise organoids for assessing drug response of CCA in vitro remains a challenge. Thus, the tissue-engineering approach might help produce CCA organoids with multi-phenotypic cells in a controllable manner. Therefore, this study aims to establish patient-derived CCA organoids using a re-assembly scheme from pre-characterized cells. From the results, explant culture was established under in vitro condition with cells grew out of CCA explants in a 2D plane. Single cells were further isolated and initially characterized with anti-cytokeratin 19 and anti-α-SMA antibodies to define the CCA cell and the cancer-associated fibroblast (CAF) populations. Further characterization of cells will be required, and drug responses of self-assembled organoids (CCA-CAF) will be elucidated in future.