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Updated: Sep 17, 2025
Standardization and Maintenance of 3D Canine Hepatic and Intestinal Organoid Cultures for Use in Biomedical Research
Published on: January 31, 2022
Canine Adrenomedullary and Pheochromocytoma Organoids: A Novel In Vitro Model
Marit F van den Berg1, Elpetra P M Timmermans-Sprang1, Fleur C Viets1
1Department Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, Utrecht 3584 CM, The Netherlands.
Context:
Given the lack of effective medical treatment for pheochromocytomas (PCCs), a reliable in vitro model is needed to explore new therapies. Organoids are three-dimensional (3D) self-renewing structures that exhibit key features of their tissue of origin, providing valuable platforms for disease modeling and drug screening.
Objective:
This study aimed to establish and characterize organoid cultures of canine normal adrenal medullas and PCCs.
Methods:
Normal adrenal medullas from healthy dogs and tumor tissue from client-owned dogs with PCC were used to develop organoids. Primary cell suspensions were cultured in a 3D matrix, and organoids were established under optimized conditions. Organoids were characterized using histology, immunohistochemistry, immunofluorescence, qPCR, and metanephrine analysis by LC-MS/MS.
Results:
Five adrenomedullary organoid lines were successfully established, demonstrating sustained growth. Organoid cultures were also derived from 9 PCCs, although expansion was limited after passages 1 to 2. Both adrenomedullary and PCC organoids expressed differentiation markers (chromogranin A, synaptophysin, phenylethanolamine N-methyltransferase) and stem/progenitor markers (nestin, SOX10). Organoids retained key functional traits, as indicated by metanephrine levels in culture supernatants, which initially mirrored primary tumor patterns. A decline in both differentiation marker expression and metanephrine levels was observed over time, possibly due to organoid dedifferentiation or selective loss of differentiated chromaffin cells.
Conclusion:
This study demonstrates the establishment of the first adrenomedullary and PCC organoid lines. While further optimization is needed, these organoids offer valuable potential as an in vitro model to investigate PCC pathophysiology and explore novel treatment strategies for this therapeutically challenging tumor.
Insights
Researchers developed the first canine organoid models from normal adrenal medullas and pheochromocytomas (PCCs). These organoids offer a promising in vitro platform for studying PCCs and testing new therapies.
Area of Science:
- Veterinary Medicine
- Oncology
- Cell Biology
Background:
- Pheochromocytomas (PCCs) lack effective treatments, necessitating reliable in vitro models for therapeutic research.
- Organoids, 3D self-renewing structures, mimic tissue characteristics, making them valuable for disease modeling and drug screening.
Purpose of the Study:
- To establish and characterize organoid cultures from canine normal adrenal medullas and PCCs.
- To evaluate the potential of these organoids as preclinical models for PCC research.
Main Methods:
- Organoids were developed from normal canine adrenal medullas and PCC tumor tissues.
- Characterization involved histology, immunohistochemistry, immunofluorescence, qPCR, and metanephrine analysis (LC-MS/MS).
Main Results:
- Successfully established five adrenomedullary and nine PCC organoid lines, with sustained growth in normal lines.
- Organoids expressed key differentiation and stem/progenitor markers, retaining functional metanephrine production mirroring primary tumors.
- Observed a decline in differentiation markers and metanephrine over time, suggesting potential dedifferentiation or cell loss.
Conclusions:
- This study reports the first canine adrenomedullary and PCC organoid lines.
- These organoids represent a valuable in vitro model for investigating PCC pathophysiology and developing novel therapeutic strategies.
