Tumoroid Culture

Learn about establishing tumoroid models for cancer research by downloading our free guidebook. This technical handbook reviews the key steps of the tumoroid workflow from sourcing tumoroids through downstream assays, and calls out key tips and tricks to help educate researchers on the nuances of tumoroid culture.

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What is a tumoroid?

Tumoroids, also known as cancer organoids, are patient-derived cancer cells grown in 3D that self-organize into multicellular structures. Compared to traditional 2D cancer cell lines, tumoroids better reflect the mutational status, gene expression levels, and phenotypes observed in patient tumors. This makes tumoroid models a vital bridge between laboratory models and clinical research. Important to keep in mind is that these are complex models, and as such, they have complex culture methods. Explore the resources below to learn more about these advanced cancer models.

How does a tumoroid compare to other cancer cell models?

2D cancer models – Traditional cancer cell model system with immortalized cancer cell lines grown as a monolayer on a culture vessel

Cancer spheroids – 3D models in which immortalized cancer cell lines are grown in suspension, forming spherical clusters

Tumoroids—(a.k.a. cancer organoids) patient-derived cancer cells grown as 3D, self-organized multicellular structures with complex culture requirements

Cancer cell models have traditionally used immortalized cancer cell lines maintained as 2D, adherent cultures. More recently, the same cells grown as 3D spheroids have been used to provide greater spatial complexity. Both systems however are limited by genetic drift and relative homogeneity that is not reflective of cancer biology. Tumoroids offer a more biologically relevant system that maintain donor-specific characteristics in long-term culture.

Why are tumoroids valuable as cancer research models?

tumoroid illustration with key values labeled

Tumoroids capture the biological complexity and relevance found in more complex cancer models, while offering scalability and cost-effectiveness compared to alternatives like animal models or organ-on-a-chip models. Tumoroids are especially valuable for complex applications, where it is important to arrive at predictive outcomes as early as possible in the research workflow.

Tumoroid culture eLearning course

The OncoPro Medium eLearning course aims to provide comprehensive training on the growth and maintenance of tumoroids using OncoPro Tumoroid Culture Medium. Accompanied by visual aids to enhance the learning experience, this interactive course is great for anyone looking to use OncoPro Medium for tumoroid culture.

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The tumoroid culture workflow

The tumoroid workflow can be divided into 5 key steps: source, culture, characterize, engineer, and assay. To get started, there are three sources for tumoroid derivation: existing tumoroid lines, tumor resections, or patient-derived xenograft (PDX) tissue. Once established, tumoroid models can then be leveraged for several applications including basic research, translational research, drug discovery, immuno-oncology, biomarker identification, and precision medicine research.

Title	Description
User Guide: OncoPro Tumoroid Culture Medium Kit	Protocols for culturing tumoroids using OncoPro Tumoroid Culture Medium in suspension and embedded culture conditions.
Protocol: Preparation of tumoroids for immunofluorescence	Workflow guidance on adapting tumoroids for immunofluorescence imaging

Application note title	Description
Tumoroid line derivation using OncoPro Tumoroid Culture Medium	Demonstrates that OncoPro Medium supports derivation of new tumoroid lines from clinical samples, such as tumor resections or patient-derived xenograft (PDX) tissues Addresses factors affecting derivation success rates (e.g., tissue quality, fresh vs frozen samples, etc.) Provides best practices for tissue handling, dissociation, and culture initiation when deriving new tumoroids
Culture of triple-negative breast cancer tumoroid lines in OncoPro Tumoroid Culture Medium	Demonstrates OncoPro Medium compatibility with triple-negative breast cancer (TNBC) tumoroids Recommended indication specific growth factors: heat-stable FGF10 and beta-estradiol Maintenance of patient-specific characteristics including morphology, growth rates, and mutational and transcriptional profiles
Enabling tumoroid-based compound screens with OncoPro Tumoroid Culture Medium	Highlights a) value of tumoroids over traditional immortalized cell lines for drug screening and b) how OncoPro suspension protocol enables high throughput workflows Addresses practical considerations such as scaling up cultures, ensuring reproducible tumoroid plating, and designing multiplexed assay readouts.

Protocols

Title	Description
User Guide: OncoPro Tumoroid Culture Medium Kit	Protocols for culturing tumoroids using OncoPro Tumoroid Culture Medium in suspension and embedded culture conditions.
Protocol: Preparation of tumoroids for immunofluorescence	Workflow guidance on adapting tumoroids for immunofluorescence imaging

Application notes

Application note title	Description
Tumoroid line derivation using OncoPro Tumoroid Culture Medium	Demonstrates that OncoPro Medium supports derivation of new tumoroid lines from clinical samples, such as tumor resections or patient-derived xenograft (PDX) tissues Addresses factors affecting derivation success rates (e.g., tissue quality, fresh vs frozen samples, etc.) Provides best practices for tissue handling, dissociation, and culture initiation when deriving new tumoroids
Culture of triple-negative breast cancer tumoroid lines in OncoPro Tumoroid Culture Medium	Demonstrates OncoPro Medium compatibility with triple-negative breast cancer (TNBC) tumoroids Recommended indication specific growth factors: heat-stable FGF10 and beta-estradiol Maintenance of patient-specific characteristics including morphology, growth rates, and mutational and transcriptional profiles
Enabling tumoroid-based compound screens with OncoPro Tumoroid Culture Medium	Highlights a) value of tumoroids over traditional immortalized cell lines for drug screening and b) how OncoPro suspension protocol enables high throughput workflows Addresses practical considerations such as scaling up cultures, ensuring reproducible tumoroid plating, and designing multiplexed assay readouts.