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Updated: Apr 4, 2026
Optimization of Renal Organoid and Organotypic Culture for Vascularization, Extended Development, and Improved Microscopy Imaging
Published on: March 28, 2020
Three Dimensional Culture of Human Renal Cell Carcinoma Organoids
Cynthia A Batchelder1, Michele L Martinez1, Nadire Duru1
1California National Primate Research Center, University of California Davis, Davis, CA, United States of America.
Abstract:
Renal cell carcinomas arise from the nephron but are heterogeneous in disease biology, clinical behavior, prognosis, and response to systemic therapy. Development of patient-specific in vitro models that efficiently and faithfully reproduce the in vivo phenotype may provide a means to develop personalized therapies for this diverse carcinoma. Studies to maintain and model tumor phenotypes in vitro were conducted with emerging three-dimensional culture techniques and natural scaffolding materials. Human renal cell carcinomas were individually characterized by histology, immunohistochemistry, and quantitative PCR to establish the characteristics of each tumor. Isolated cells were cultured on renal extracellular matrix and compared to a novel polysaccharide scaffold to assess cell-scaffold interactions, development of organoids, and maintenance of gene expression signatures over time in culture. Renal cell carcinomas cultured on renal extracellular matrix repopulated tubules or vessel lumens in renal pyramids and medullary rays, but cells were not observed in glomeruli or outer cortical regions of the scaffold. In the polysaccharide scaffold, renal cell carcinomas formed aggregates that were loosely attached to the scaffold or free-floating within the matrix. Molecular analysis of cell-scaffold constructs including immunohistochemistry and quantitative PCR demonstrated that individual tumor phenotypes could be sustained for up to 21 days in culture on both scaffolds, and in comparison to outcomes in two-dimensional monolayer cultures. The use of three-dimensional scaffolds to engineer a personalized in vitro renal cell carcinoma model provides opportunities to advance understanding of this disease.
Insights
Researchers developed 3D models of renal cell carcinoma (RCC) using natural scaffolds. These models successfully maintained individual tumor characteristics for up to 21 days, offering a promising tool for personalized cancer therapy research.
Area of Science:
- Oncology
- Biomaterials Science
- Regenerative Medicine
Background:
- Renal cell carcinoma (RCC) is a heterogeneous cancer with diverse clinical behaviors and treatment responses.
- Developing patient-specific in vitro models is crucial for advancing personalized therapies for RCC.
- Existing 2D cell cultures often fail to replicate the complex in vivo tumor microenvironment.
Purpose of the Study:
- To engineer patient-specific in vitro models of renal cell carcinoma (RCC) using three-dimensional (3D) culture techniques.
- To evaluate the efficacy of natural scaffolding materials, specifically renal extracellular matrix and a novel polysaccharide scaffold, in maintaining tumor phenotypes.
- To assess the potential of these 3D models for personalized therapy development in RCC.
Main Methods:
- Human renal cell carcinomas were characterized using histology, immunohistochemistry, and quantitative PCR.
- Tumor cells were cultured on renal extracellular matrix and a novel polysaccharide scaffold in 3D.
- Cell-scaffold interactions, organoid formation, and gene expression signatures were analyzed over 21 days.
- Comparisons were made to 2D monolayer cultures.
Main Results:
- RCC cells cultured on renal extracellular matrix repopulated tubular and vascular structures within the scaffold.
- In the polysaccharide scaffold, RCC cells formed aggregates with loose attachment or remained free-floating.
- Both 3D scaffolds sustained individual tumor phenotypes, confirmed by molecular analysis (immunohistochemistry, qPCR), for up to 21 days.
- 3D cultures demonstrated superior maintenance of tumor characteristics compared to 2D cultures.
Conclusions:
- Three-dimensional scaffolds can be used to engineer personalized in vitro models of renal cell carcinoma.
- These models effectively maintain distinct tumor phenotypes, offering a platform for studying disease biology.
- The developed 3D models hold promise for advancing the understanding and personalized treatment of renal cell carcinoma.
