Cancer research has always been challenging due to the complexity and heterogeneity of tumors. Traditional two-dimensional (2D) cell cultures and animal models have been the backbone of cancer research, but they come with significant limitations.

Introducing tumoroids—three-dimensional (3D) cell culture models that replicate the tumor microenvironment more accurately. They are essentially organoids that resemble tumors derived from patient tumor cells. They are now instrumental in drug screening and studying the dynamic growth and behavior of cancer cells in laboratory settings.

Here, we highlight the general features, mechanisms, applications, challenges, and future projections of this cutting-edge technology that is redefining cancer research.

General features of tumoroids

Tumoroids are 3D structures grown in vitro from primary tumor cells harvested from cancer patients. They can replicate the architecture and microenvironment of the original tumor¹. Unlike 2D cultures, tumoroids preserve the natural cell morphology, gene expression, and cell-cell interactions.
This physiologically relevant model is ideal for elucidating vital processes like cancer formation, growth, and metastasis by monitoring the primary influencing factors. These features grant tumoroids a sensitive and valid tool for exploring tumor biology, drug response, and resistance mechanisms, posing significant advantages over conventional approaches². Moreover, it helps reduce the need for human and animal testing, making cancer research more ethical and sustainable³.

Organoids vs tumoroids

While both organoids and tumoroids are 3D cell culture models, they fulfill different purposes and have distinct characteristics. Organoids are derived from stem cells and can differentiate into multiple cell types to form mini-organs. They are applied to study normal organ development and related diseases. Tumoroids, on the other hand, are derived from tumor cells and are used exclusively to model cancer. They preserve the genetic and phenotypic features of the original tumor, making them pivotal for cancer research⁴.

Mechanisms related to tumoroid formation

A tumor is a complex assembly of cancer cells, host cells, secreted factors, and the extracellular matrix (ECM). Tumor cells cause substantial changes in the surrounding tissues to facilitate growth and progression. The tumor microenvironment (TME) is continually evolving and varies among tumor types, typically including immune cells, stromal cells, blood vessels, and the ECM⁵.
Given the intricate nature of the TME and the roles of various cell types, a molecular network comprising multiple steps is involved in tumoroid formation². Firstly, tumor cells are isolated from patient tissue and then cultured in a 3D matrix, such as Matrigel, which supports cell growth and organization. These cells then proliferate and self-organize into structures that mimic the original tumor. The ECM and different growth factors are crucial in this process, influencing cell behavior and interactions⁶.

Tumoroid culture workflow

The workflow for culturing tumoroids involves a few key steps:
Sourcing Tumoroids: Tumoroids can be obtained through surgical resections or biopsies from patient tumor samples. These samples are then processed to isolate tumor cells cultured in a 3D matrix⁷.

Culturing Techniques: The culturing of tumoroids involves media preparation, cell seeding, and maintenance. The cells are embedded in a matrix and cultured in specialized media that supports their growth and differentiation. Regular monitoring and maintenance are required to ensure the health and viability of the tumoroids⁸.

Characterization: Characterizing tumoroids entails evaluating their morphology, genetic and phenotypic profiles, and functional properties. Techniques such as immunohistochemistry, gene expression analysis, and drug sensitivity assays are commonly used to assess the quality and properties of cultured tumoroids⁹.

Advantages and applications

Tumoroids provide a more accurate model of human tumors than traditional methods, which benefits several key applications¹⁰.

Accurate cancer modeling: As mentioned above, tumoroids can faithfully replicate the structure, function, and genetic makeup of primary tumors, bridging the gap between 2D cell cultures and in vivo models.

Therapeutic response prediction: Tumoroids can be employed to predict how individual patients will respond to specific treatments, aiding in the development of personalized treatment plans.

Drug discovery and testing: They function as valuable models for screening potential therapeutic compounds and identifying new drug targets, an imperative step in drug discovery.

Investigating resistance mechanisms: Tumoroids help researchers understand why specific cancers become resistant to treatments, resulting in the development of more effective therapies.

Modeling tumor microenvironment: Tumoroids can be used to study the interactions between cancer cells and their surrounding environment, including immune cells and blood vessels, that play major roles in dictating the metastatic potential of the disseminating cells¹¹.

Optimizing treatment strategies: By providing a more accurate model of human tumors, tumoroids can help refine existing treatment strategies and explore new therapeutic approaches.

Challenges and future prospects

Despite their advantages, tumoroids also face several challenges. Standardized protocols are urgently needed to ensure reproducibility across different labs². Also, large-scale production of tumoroids remains an immense complication, limiting their use in high-throughput screening, a central stage in drug discovery and development⁴. Moreover, tumoroid cultures are complex and require very specific expertise and specialized equipment, such as bioreactors and microfluidic devices, which can be costly and difficult to manage¹².

Looking ahead, the future of tumoroids in cancer research is rather promising. Advances in bioprinting and microfluidics could enhance the complexity and scalability of tumoroid models. Additionally, integrating tumoroids with other technologies, such as single-cell sequencing and CRISPR, could provide deeper insights into tumor biology and lead to the development of more efficacious treatments¹³.

Tumoroids represent a solid progress in cancer research, offering a more accurate and physiologically relevant model for studying tumors and testing new treatments. Collaborative communication among cell biologists, materials scientists, and engineers is essential to develop scaffold-based 3D cell culturing systems that can effectively address increasing biological and technical trials. While challenges persist, active research and scientific advancements hold the promise of overcoming these obstacles and unlocking the full potential of tumoroids in the battle against cancer.

References

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