[Establishment and identification of organoid derived from patients with pancreatic cancer based on suspension culture]

Abstract

Objective: To explore the application of the suspension culture method in pancreatic cancer organoid construction. Methods: Cell suspensions obtained from 8 pancreatic cancer tissue samples at the Second Hospital of Lanzhou University between July 2023 and March 2024, were prepared by digested pancreatic cancer tumor tissues using mixed enzymes, inoculated into ultra-low adsorption culture plates for suspension culture, and when the organoids were cultured to a certain size, passaging and freezing were initiated, and their structural morphology was observed by inverted microscope. Hematoxylin-eosin (HE) staining showed that pancreatic cancer organoids were lumpy or irregularly tubular, with obvious nuclear atypia, and were remarkably similar in tissue structure to pancreatic cancer tissue. Results: Among the 8 pancreatic cancer tissue samples, pancreatic cancer organoids were successfully constructed in three patients, and HE staining showed that pancreatic cancer organoids had a high degree of structural similarity with tumor tissues. Immunohistochemistry suggested that CK7, CK19, P53, and Ki-67 were expressed in the pancreatic cancer organoids and tumor tissues of case origin in more or less the same way. Conclusion: The suspension culture method is able to construct pancreatic cancer-like organs that are approximately the same as the originating tumor tissues at the tissue level.