[Study on the establishment, assessment, and drug sensitivity testing for intrahepatic cholangiocarcinoma organoid models based on the suspension culture method].

Objective: To investigate the feasibility of establishing intrahepatic cholangiocarcinoma (ICC) organoid models based on the suspension culture method. Methods: Fresh tumor tissue specimens were prepared from six patients with intrahepatic cholangiocarcinoma using digestive enzymes for a cell suspension. The cells were cultured as organoids in ultra-low attachment plates. Passaging and cryopreservation were used once the organoids reached an appropriate size. The formation of the organoids was monitored and recorded using an inverted microscope during the culture process. The organoids were collected and detected by hematoxylin-eosin staining and immunohistochemistry. The protein expressional conditions were examined. Results: Stable passaged and cryopreserved organoids from three tumor tissue samples were successfully constructed. Hematoxylin-eosin staining results showed that the organoids were structurally similar to the primary tissue. Immunohistochemical staining results demonstrated that CK7, CK19, E-cadherin, and vimentin were all positively expressed, with Ki67 positivity rates of approximately 50%, 30%, and 80%, respectively. Conclusion: The suspension culture method can construct organoid models in terms of tissue structure and protein expression that exhibit tumor characteristics of intrahepatic cholangiocarcinoma.