Related Experiment Video
Updated: Aug 5, 2026
Establishment of Pancreatic Cancer-Derived Tumor Organoids and Fibroblasts From Fresh Tissue
Published on: May 26, 2023
Comparative evaluation of chemotherapy and CAR-T cell therapy in genetically engineered self-sustaining primary
Xu Wang1, Ruijie Wang2, Xuan Lin1
1Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, No. 270 Dong an Road, Xu-Hui District, Shanghai, 200032, China; Department of Oncology, Shanghai Medical College, Fudan University, No.270 Dong an Road, Xu-Hui District, Shanghai, 200032, China; Shanghai Pancreatic Cancer Institute, No.270 Dong an Road, Xu-Hui District, Shanghai, 200032, China; Pancreatic Cancer Institute, Fudan University, No.270 Dong an Road, Xu-Hui District, Shanghai, 200032, China.
Background:
Broader research and translational use of patient-derived organoids (PDOs) in pancreatic ductal adenocarcinoma (PDAC) remain constrained by complex and costly culture requirements. Here, we developed a conditional transgenic strategy to generate self-sustaining primary PDAC organoids and explored its feasibility for therapeutic evaluation.
Methods:
We established a proof-of-concept PDAC organoid biobank (n = 10). Using a doxycycline-inducible lentiviral Tet-On system, we engineered organoids to express essential cytokines RSPO1 and WNT3A, enabling growth in supplement-depleted media. We then performed exploratory assessments of FOLFIRINOX-mimetic chemotherapy and EPHA2-targeted CAR-T cells in transgenic organoids, together with histopathological and clinicopathological co-analysis.
Results:
Doxycycline-induced RSPO1/WNT3A expression rescued the impaired growth and passaging caused by exogenous cytokine depletion. In simplified media, transgenic organoids retained patient-specific histological and molecular features. Sensitivity to FOLFIRINOX-mimetic treatment and EPHA2 CAR-T cells varied among different patients, and the in vitro responses showed exploratory concordance with the corresponding clinical courses in a small subset.
Conclusion:
This conditional transgenic approach reduces culture complexity while preserving key phenotypic and functional characteristics of PDAC organoids, providing a feasible proof-of-concept platform for exploratory evaluation of chemotherapy and immunotherapy responses.
Insights
A new transgenic method simplifies culturing patient-derived pancreatic cancer organoids. This approach enables easier study of chemotherapy and immunotherapy responses in pancreatic ductal adenocarcinoma.
Area of Science:
- Oncology
- Stem Cell Biology
- Translational Medicine
Background:
- Patient-derived organoids (PDOs) are valuable for pancreatic ductal adenocarcinoma (PDAC) research but face challenges due to complex culture needs.
- Current methods limit broader research and therapeutic applications of PDAC organoids.
- Developing simplified organoid culture systems is crucial for advancing PDAC studies.
Purpose of the Study:
- To develop a simplified, self-sustaining culture system for primary pancreatic ductal adenocarcinoma organoids.
- To create a proof-of-concept platform for evaluating therapeutic responses in PDAC organoids.
- To reduce the complexity and cost associated with PDAC organoid culture.
Main Methods:
- Established a biobank of 10 PDAC organoids.
- Engineered organoids using a doxycycline-inducible Tet-On system to express RSPO1 and WNT3A cytokines.
- Assessed organoid response to FOLFIRINOX-mimetic chemotherapy and EPHA2-targeted CAR-T cells in simplified media.
Main Results:
- Doxycycline induction successfully enabled organoid growth in cytokine-depleted media.
- Transgenic organoids maintained patient-specific histological and molecular features in simplified conditions.
- Observed variable in vitro sensitivity to chemotherapy and CAR-T cells, with exploratory concordance to clinical outcomes.
Conclusions:
- A conditional transgenic strategy simplifies PDAC organoid culture while preserving key characteristics.
- This approach offers a feasible platform for exploring chemotherapy and immunotherapy efficacy.
- The developed method facilitates translational research and therapeutic evaluation in pancreatic cancer.
