Pancreatic duct-like organoids (PDLOs) are being increasingly used as a model system for investigating the pathology of pancreatic diseases, evaluating new drug treatments, and enabling the advancement of cell and gene therapy research. In this study, we demonstrate the ability to culture and characterize human iPSC-derived PDLOs using
R&D Systems reagents and analysis platforms. The PDLOs that were generated expressed high levels of mature pancreatic ductal cell markers, providing clear evidence that the differentiation was efficient. Further analysis by immunofluorescent staining and a CFTR activity assay showed that the PDLOs had a polarized epithelial layer and functional CFTR ion channels, confirming that this organoid system accurately models the structure and function of normal pancreatic ductal tissue.
Key Takeaways
- Flow cytometry analysis demonstrated that 95% of the cells in the final PDLO cell population expressed KRT19 and 40% expressed SOX9, two characteristic markers of pancreatic ductal cells.
- Orthogonal validation using Simple Western™ technology confirmed the expression of the NKX6.1 and PDX-1 progenitor markers decreased during differentiation, while the mature ductal cell markers, SOX9, KRT19, and E-Cadherin were expressed at high levels in the PDLOs.
- Immunofluorescent staining showed the ductal cell markers, KRT19, SOX9, and Claudin-1 were expressed in the expected locations and confirmed the high quality of the organoid structures.
- The PDLOs were shown to have functional CFTR ion channels and treatment with CFTR inhibitors suggested they could serve as an effective drug screening platform for testing the efficacy of drugs aimed at treating pancreatic diseases.