Gibco OncoPro Tumoroid Culture Medium streamlines the culture of patient-derived cancer organoids (tumoroids) to help make physiologically relevant cancer models more accessible. This adaptable culture system is readily integrated into your workflow so you can upgrade your media and maintain your results. Overall, OncoPro medium offers several key benefits to cancer researchers:

Cancer icon

Optimized—OncoPro medium was developed to support expansion of tumoroids without overgrowth of non-malignant cells. As such, this culture system helps ensure preservation of donor-specific characteristics, including mutational profiles and gene expression patterns, in long-term culture.

 Gibco bottle icon

Streamlined—OncoPro medium is a modular media system, consisting of four components that can be supplemented with indication-specific growth factors as needed. Its suspension culture capability introduces a simplified tumoroid culture workflow, helping save time and resources.

Split arrow icon

Adaptable—OncoPro medium is compatible with a suspension or embedded tumoroid workflow, supports scale-up and automation, and sustains tumoroids across multiple cancer indications. OncoPro medium is also compatible with traditional applications and assays.


Customer stories

Erik Goka

"The Gibco OncoPro Tumoroid Culture Medium system allows us to seamlessly integrate tumor organoids into our research and drug discovery processes. With the tumoroid media system we can propagate all our organoid models in the same culture medium system irrespective of the tissue of origin. This media system has been instrumental in allowing us to bring tumor organoids into our lab for everyday use."

Erik Goka, PhD
Vice President of Biology
Revere Pharmaceuticals

Shawn Fahl

"The OncoPro media has greatly accelerated our development of long-term primary tumor culture systems. The media is easy to make, with a minimal amount of separate components compared to other systems, and the protocols are straightforward, allowing the transition from culture initiation to viable long-term cultures to be seamless and quick."

Shawn Fahl, PhD
V.P. Lab Operations, Cell Services & R&D,
BiospecimensDiscovery Life Sciences


Benefits and performance data of OncoPro medium

Discover how OncoPro medium was optimized for tumoroid culture

OncoPro Tumoroid Culture Medium is a novel media system that was specifically developed for the expansion of patient-derived tumoroids. Over 40 conditions were tested during the development of OncoPro to identify the combination of basal media, supplements, and growth factors that would best support long-term maintenance of donor-specific characteristics, including mutational profiles and gene expression patterns.

Figure 1. Design of experiment studies were used to fully optimize the OncoPro medium formulation for tumoroid growth. Combinations of medium components were tested in designed experiments to elucidate main effects and interactions of additives on tumoroid growth. In this example, two tumoroid lines were tested using 25 media recipes, with 1-2 replicate wells per condition. (A) Heat map of number of viable cells detected per media condition as measured by PrestoBlue HS assay. (B) Images of tumoroids in medium optimization experiments. Different conditions were tested on dissociated cells seeded from an identical initial cell pool, and conditions advantageous to tumoroid growth and retention of genetic and phenotypic characteristics were identified. (C) Design of experiment output for tumoroid line growth response to media optimization experimental conditions. Figure adapted from JMP 16 software output.

Figure 2. OncoPro medium selectively expands cancer cells. (A) Marker expression (flow cytometry) of cells freshly isolated from a colorectal tumor resection (never in culture) and tumoroid cells after line establishment (i.e., 7 passages in OncoPro medium). The panel included markers for epithelial-derived cells (EpCAM), colorectal cancer cells (CEACAM), immune cells (CD45), endothelial cells (CD31), and mesenchymal cells (vimentin). Data reveal preferential outgrowth of epithelial colorectal cancer cells over immune or endothelial cells. (B) Representative immunofluorescence image of EpCAM and actin (phalloidin) expression demonstrate that tumoroids are highly enriched for EpCAM-positive tumor cells.

Figure 3. OncoPro medium helps maintain mutational profiles of tumoroid lines over long-term culture. Correlation of germline and somatic single nucleotide variations (SNVs) between early and later-passage tumoroid cultures. Each dot represents the variant allelic frequency (VAF) for one genetic locus covered by the Oncomine Comprehensive Assay v3. Figure adapted from JMP 16 software output.

Understand how OncoPro medium streamlines media formulation and tumoroid culture

OncoPro medium affords researchers workflow versatility by letting users decide if they want to utilize established embedded culture methods or transition to suspension methods to help save both time and resources.

Scientific diagrams showing tumoroid culture methods

Figure 4. The OncoPro medium suspension culture method greatly simplifies tumoroid passaging, reducing hands-on time from 140 minutes for embedded culture to 60 minutes for suspension culture. (A) Schematic of culture method steps for passaging tumoroids in embedded culture. Boxes highlight the pain points of disrupting extracellular matrix (ECM) domes, losing cells trapped in ECM during washing, and pipetting and waiting for ECM domes to solidify, respectively. (B) Schematic of culture method steps for passaging tumoroids in suspension culture, which addresses these pain points due to reduced ECM requirement.
Figure created with BioRender.com.

Table comparing suspension vs embedded culture

Figure 5. Tumoroid cultures are readily scalable with the OncoPro medium suspension culture method. The number and format of culture vessels required to obtain increasing scale of tumoroid cultures are more user- and resource-friendly in suspension culture when compared to traditional embedded culture methods. (A) Suspension methods allow for ready expansion of 100 million cells within one passage, whereas the number of basement membrane extract (BME) domes required in embedded culture would be prohibitive for many users. (B) Average amount of BME required to harvest 100 million cells for 9 tumoroid lines using different culture methods. On average, suspension culture required less BME than embedded culture (mean ± SEM, not significant).

Figure 6. Patient-specific gene expression profiles are highly correlated between samples cultured using the streamlined suspension method and those cultured using traditional embedded methods. Ward hierarchical clustering and Spearman’s ρ correlations of gene expression for a panel of 1423 cancer-related genes across a variety of tumoroid lines representing four different cancer indications. Gene expression assayed by Ion AmpliSeq Transcriptome Human Gene Expression Panel, Chef-Ready Kit. Figures adapted from JMP 16 software output. With the exception of colorectal tumoroid line Z, established tumoroid models provided by NCI PDMR. We thank the PDMR and their contributing institutions for their contributions to this work.

Learn how OncoPro medium functions as an adaptable system

OncoPro Tumoroid Culture Medium Kit is a flexible system designed to address complex research needs. OncoPro medium accommodates multiple cancer indications using a single media kit and the scalable and automation-friendly suspension workflow facilitates smooth transitions to various downstream assays. The absence of Wnt, R-spondin, Noggin, and small molecule inhibitors make OncoPro well-suited for advanced biological studies and screening applications.

Figure 7. OncoPro medium is compatible with four different cancer indications, for both suspension and embedded culture. Morphology of NCI PDMR tumoroid lines expanded in NCI PDMR homebrew media (embedded culture), or in OncoPro medium embedded or suspension culture. Scale bar = 400 µm. Established tumoroid models provided by NCI PDMR. We thank the PDMR and their contributing institutions for their contributions to this work.

Figure 8. OncoPro medium is compatible with four different cancer indications, for both suspension and embedded culture. Cumulative population doublings of cells expanded in OncoPro medium suspension culture (purple), OncoPro medium embedded culture (red), or alternative media system (media system in which the line was derived prior to procurement) embedded culture (green). In general, tumoroids expanded in each culture condition with comparable growth rates. With the exception of colorectal tumoroid line Z, established tumoroid models provided by NCI PDMR. We thank the PDMR and their contributing institutions for their contributions to this work.

Figure 9. OncoPro medium helps maintain patient-specific gene expression profiles from lines derived in alternative media and embedded culture. Principal component analysis (PCA) plot comparing gene expression levels across tumoroid lines and culture conditions. Tumoroid lines were expanded in parallel in multiple media and culture conditions for 2-7 passages post-thaw prior to quantification of expression levels across over >20,000 human RefSeq genes using the Ion AmpliSeq Transcriptome Human Gene Expression Kit. Tumoroid lines tested were 782815-120-R-V1-organoid (Colorectal A), 919269-233-R3-V2-organoid (Colorectal B), 349418-098-R-V2-organoid (Lung), 549293-155-R-V1-organoid (Head and Neck), and 699152-130-R-V1-organoid (Pancreas). With the exception of colorectal tumoroid line Z, established tumoroid models were provided by NCI PDMR. We thank the PDMR and their contributing institutions for their contributions to this work. Figure adapted from TAC 4.0 software output.

Discover how OncoPro medium was optimized for tumoroid culture

OncoPro Tumoroid Culture Medium is a novel media system that was specifically developed for the expansion of patient-derived tumoroids. Over 40 conditions were tested during the development of OncoPro to identify the combination of basal media, supplements, and growth factors that would best support long-term maintenance of donor-specific characteristics, including mutational profiles and gene expression patterns.

Figure 1. Design of experiment studies were used to fully optimize the OncoPro medium formulation for tumoroid growth. Combinations of medium components were tested in designed experiments to elucidate main effects and interactions of additives on tumoroid growth. In this example, two tumoroid lines were tested using 25 media recipes, with 1-2 replicate wells per condition. (A) Heat map of number of viable cells detected per media condition as measured by PrestoBlue HS assay. (B) Images of tumoroids in medium optimization experiments. Different conditions were tested on dissociated cells seeded from an identical initial cell pool, and conditions advantageous to tumoroid growth and retention of genetic and phenotypic characteristics were identified. (C) Design of experiment output for tumoroid line growth response to media optimization experimental conditions. Figure adapted from JMP 16 software output.

Figure 2. OncoPro medium selectively expands cancer cells. (A) Marker expression (flow cytometry) of cells freshly isolated from a colorectal tumor resection (never in culture) and tumoroid cells after line establishment (i.e., 7 passages in OncoPro medium). The panel included markers for epithelial-derived cells (EpCAM), colorectal cancer cells (CEACAM), immune cells (CD45), endothelial cells (CD31), and mesenchymal cells (vimentin). Data reveal preferential outgrowth of epithelial colorectal cancer cells over immune or endothelial cells. (B) Representative immunofluorescence image of EpCAM and actin (phalloidin) expression demonstrate that tumoroids are highly enriched for EpCAM-positive tumor cells.

Figure 3. OncoPro medium helps maintain mutational profiles of tumoroid lines over long-term culture. Correlation of germline and somatic single nucleotide variations (SNVs) between early and later-passage tumoroid cultures. Each dot represents the variant allelic frequency (VAF) for one genetic locus covered by the Oncomine Comprehensive Assay v3. Figure adapted from JMP 16 software output.

Understand how OncoPro medium streamlines media formulation and tumoroid culture

OncoPro medium affords researchers workflow versatility by letting users decide if they want to utilize established embedded culture methods or transition to suspension methods to help save both time and resources.

Scientific diagrams showing tumoroid culture methods

Figure 4. The OncoPro medium suspension culture method greatly simplifies tumoroid passaging, reducing hands-on time from 140 minutes for embedded culture to 60 minutes for suspension culture. (A) Schematic of culture method steps for passaging tumoroids in embedded culture. Boxes highlight the pain points of disrupting extracellular matrix (ECM) domes, losing cells trapped in ECM during washing, and pipetting and waiting for ECM domes to solidify, respectively. (B) Schematic of culture method steps for passaging tumoroids in suspension culture, which addresses these pain points due to reduced ECM requirement.
Figure created with BioRender.com.

Table comparing suspension vs embedded culture

Figure 5. Tumoroid cultures are readily scalable with the OncoPro medium suspension culture method. The number and format of culture vessels required to obtain increasing scale of tumoroid cultures are more user- and resource-friendly in suspension culture when compared to traditional embedded culture methods. (A) Suspension methods allow for ready expansion of 100 million cells within one passage, whereas the number of basement membrane extract (BME) domes required in embedded culture would be prohibitive for many users. (B) Average amount of BME required to harvest 100 million cells for 9 tumoroid lines using different culture methods. On average, suspension culture required less BME than embedded culture (mean ± SEM, not significant).

Figure 6. Patient-specific gene expression profiles are highly correlated between samples cultured using the streamlined suspension method and those cultured using traditional embedded methods. Ward hierarchical clustering and Spearman’s ρ correlations of gene expression for a panel of 1423 cancer-related genes across a variety of tumoroid lines representing four different cancer indications. Gene expression assayed by Ion AmpliSeq Transcriptome Human Gene Expression Panel, Chef-Ready Kit. Figures adapted from JMP 16 software output. With the exception of colorectal tumoroid line Z, established tumoroid models provided by NCI PDMR. We thank the PDMR and their contributing institutions for their contributions to this work.

Learn how OncoPro medium functions as an adaptable system

OncoPro Tumoroid Culture Medium Kit is a flexible system designed to address complex research needs. OncoPro medium accommodates multiple cancer indications using a single media kit and the scalable and automation-friendly suspension workflow facilitates smooth transitions to various downstream assays. The absence of Wnt, R-spondin, Noggin, and small molecule inhibitors make OncoPro well-suited for advanced biological studies and screening applications.

Figure 7. OncoPro medium is compatible with four different cancer indications, for both suspension and embedded culture. Morphology of NCI PDMR tumoroid lines expanded in NCI PDMR homebrew media (embedded culture), or in OncoPro medium embedded or suspension culture. Scale bar = 400 µm. Established tumoroid models provided by NCI PDMR. We thank the PDMR and their contributing institutions for their contributions to this work.

Figure 8. OncoPro medium is compatible with four different cancer indications, for both suspension and embedded culture. Cumulative population doublings of cells expanded in OncoPro medium suspension culture (purple), OncoPro medium embedded culture (red), or alternative media system (media system in which the line was derived prior to procurement) embedded culture (green). In general, tumoroids expanded in each culture condition with comparable growth rates. With the exception of colorectal tumoroid line Z, established tumoroid models provided by NCI PDMR. We thank the PDMR and their contributing institutions for their contributions to this work.

Figure 9. OncoPro medium helps maintain patient-specific gene expression profiles from lines derived in alternative media and embedded culture. Principal component analysis (PCA) plot comparing gene expression levels across tumoroid lines and culture conditions. Tumoroid lines were expanded in parallel in multiple media and culture conditions for 2-7 passages post-thaw prior to quantification of expression levels across over >20,000 human RefSeq genes using the Ion AmpliSeq Transcriptome Human Gene Expression Kit. Tumoroid lines tested were 782815-120-R-V1-organoid (Colorectal A), 919269-233-R3-V2-organoid (Colorectal B), 349418-098-R-V2-organoid (Lung), 549293-155-R-V1-organoid (Head and Neck), and 699152-130-R-V1-organoid (Pancreas). With the exception of colorectal tumoroid line Z, established tumoroid models were provided by NCI PDMR. We thank the PDMR and their contributing institutions for their contributions to this work. Figure adapted from TAC 4.0 software output.

Looking for more information about OncoPro Tumoroid Culture Medium or how you can request a sample quote?

Contact us


Explore technical literature demonstrating the performance and application of OncoPro Tumoroid Culture Medium

Application note titleDescription
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.

Access five unique posters from our research and development scientists that highlight the key features of OncoPro medium and how it can be applied in certain tumoroid culture workflows such as drug screening and immuno-oncology. 

Poster TitleDescription

Derivation & long-term maintenance of patient-derived tumoroid lines in a defined, serum-free medium

  • Maintenance of growth rates, patient phenotype and genotype for ≥40 passages
  • Support for multiple cancer indications using tissue-specific growth factors
  • Scale up to >1 billion cancer organoid cells with easy-to-use suspension method

Expansion of established patient-derived tumoroids in a novel serum-free, Wnt agonist-free media system

  • Long-term maintenance of mutational status and gene expression levels in both embedded and suspension formats
  • Wnt-related signaling pathways are not differentially activated following culture in Wnt agonist-free OncoPro Tumoroid Culture Medium

How low can you go: Maintenance of tumoroid phenotype with a highly scalable and automation-compatible reduced-ECM suspension culture method

  • Suspension culture using OncoPro medium is compatible with existing tumoroid lines
  • Patient phenotype and genotype in suspension conditions using OncoPro medium are comparable with embedded method

Multiplexed plate-reader based drug screening of 3D-tumoroid models

  • Development and optimization of drug screening assay using tumoroids
  • Drug response comparison of tumoroids and immortalized cell lines 2D and 3D
  • Compound screening application using tumoroids
Engineering patient-derived tumoroids to enable high-throughput screening:Immuno-oncology workflows
  • Establishment of GFP-expressing tumoroid reporter pool
  • Demonstration of NK-cell mediated cytotoxicity against tumoroids
  • Comparison of primary, iPSC-derived, and NK lines cytotoxicity
Application note titleDescription
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.

Access five unique posters from our research and development scientists that highlight the key features of OncoPro medium and how it can be applied in certain tumoroid culture workflows such as drug screening and immuno-oncology. 

Poster TitleDescription

Derivation & long-term maintenance of patient-derived tumoroid lines in a defined, serum-free medium

  • Maintenance of growth rates, patient phenotype and genotype for ≥40 passages
  • Support for multiple cancer indications using tissue-specific growth factors
  • Scale up to >1 billion cancer organoid cells with easy-to-use suspension method

Expansion of established patient-derived tumoroids in a novel serum-free, Wnt agonist-free media system

  • Long-term maintenance of mutational status and gene expression levels in both embedded and suspension formats
  • Wnt-related signaling pathways are not differentially activated following culture in Wnt agonist-free OncoPro Tumoroid Culture Medium

How low can you go: Maintenance of tumoroid phenotype with a highly scalable and automation-compatible reduced-ECM suspension culture method

  • Suspension culture using OncoPro medium is compatible with existing tumoroid lines
  • Patient phenotype and genotype in suspension conditions using OncoPro medium are comparable with embedded method

Multiplexed plate-reader based drug screening of 3D-tumoroid models

  • Development and optimization of drug screening assay using tumoroids
  • Drug response comparison of tumoroids and immortalized cell lines 2D and 3D
  • Compound screening application using tumoroids
Engineering patient-derived tumoroids to enable high-throughput screening:Immuno-oncology workflows
  • Establishment of GFP-expressing tumoroid reporter pool
  • Demonstration of NK-cell mediated cytotoxicity against tumoroids
  • Comparison of primary, iPSC-derived, and NK lines cytotoxicity


Order your OncoPro Tumoroid Culture Medium Kit

OncoPro product photo

OncoPro Tumoroid Culture Medium recommended reagents

The reagents below are recommended for culturing cancer organoids using OncoPro Tumoroid Culture Medium Kit.

We have demonstrated the OncoPro Tumoroid Culture Medium Kit supports the culture of several epithelial-derived tumoroids, but additional growth factors may be required for some indications.

For other indications, please Contact our support team to learn how you may be able to use OncoPro medium.

The following recommended reagents can be purchased directly from the Fisher Scientific catalog: 

Recommended reagents from Thermo Fisher Scientific

Thermo Fisher Scientific also offers Gibco OncoPro Tumoroid Cell Lines for purchase. Utilize our general inquiry form below to get a quote and additional information.

Submit an inquiry


Engage with additional resources for tumoroid culture


Discover other products to support culture of tumoroids

Cell culture plastics family image

Nunc Cell Culture Plastics

Thermo Scientific Nunc cell culture plastics offer non-treated flasks to support tumoroid culture.

A bottle of HPLM with a graphic rendering of a cell

Gibco HPLM

Human plasma-like medium is a physiologically relevant medium useful for downstream assays.



For Research Use Only. Not for use in diagnostic procedures.