Establishment and characterization of primary cell culture of urothelial carcinoma: A platform for functional study in cancer

Purpose: It has been known that tumor heterogeneity commonly exists at both the intertumoral and intratumoral level. This kind of complexity causes the difficulties to find a solution for the disease using single modality. Here, we want to collect and create a Taiwanese urological cancer database, establish primary cell line culture from surgical specimens, and find their potential application for pharmacological therapy.

Materials and Methods: Normal and pathological tissues from urological cancers were collected immediately after the specimens were extracted. The surgical specimens were minced with sterile scissors and cultured in tissue plate. The normal tissues were also collected and stored in liquid nitrogen. In order to verify the consistency of the tumor tissue, normal tissue and derived primary cell, we used STR assay to perform this experiment. We also used phenotypical analysis to characterize the primary cell lines we have established. Besides, we established cancer xenografts from clinical materials by using mouse models for in vivo study of human urological carcinomas.

Results: We have successfully established the primary cell cultures from different tumor grade and tumor type. In order to characterize the primary cells we get, we used cytofluorometrical analysis to perform this experiment. All the cells stained with desmin were negative, confirming that the cultured cells were not contaminated by fibroblast or muscle tissue. In the STR assay, the results showed that almost all tissues and primary cell lines still maintained their consistency even after passaging. We also set up the patient-derived xenograft (PDX) model in the urothelial carcinomas.

Conclusion: This is a combination of basic and translation research. By setting up a good platform in urothelial carcinoma cell lines, our efforts may help us to advance our understanding of the urological cancers and develop many unique cancer cell lines for functional research in cancers.