Protocol for generating a pancreatic cancer organoid associated with heterogeneous tumor microenvironment.

Abstract

Pancreatic ductal adenocarcinoma (PDAC) organoids that simulate the tumor microenvironment (TME) are an effective tool to identify how TME affects PDAC malignancy. We present a protocol for generating a fused pancreatic cancer organoid (FPCO) that partly reproduces the TME, including heterogeneous cancer-associated fibroblasts (CAFs), using patient-derived PDAC cells and human-induced pluripotent cell-derived endothelial and mesenchymal cells. We also describe the procedure for analyzing FPCO characteristics. FPCO can provide a platform for establishing a reliable drug screening system. For complete details on the use and execution of this protocol, please refer to Takeuchi et al.¹.