Cultivating Ex Vivo Patient-Derived Glioma Organoids Using a Tissue Chopper
January 19th, 2024
This study introduces an automated method to generate patient-derived glioblastoma 3-dimensional organoids utilizing a tissue chopper. The method provides a suitable and effective approach to obtain such organoids for therapeutical testing.
So our research focuses on improving generation of patient derived organoids for glioblastoma patients. And our key questions here revolve around whether this approach can efficiently produce more organoids, reduce manufacturing time, and at the same time maintain the characteristics of the original tumor. And we would use that for an effective drug and immunotherapy screening in glioblastoma patients.
The recent developments in the glioblastoma field include advancement in understanding the genetic and molecular aspects leading to the recognition of new targets. Additionally, there's a great growing emphasis on using 3D organoids, to better replicate the complexity of tumors and allow drug testing. Currently, technologies like manual dissection, enzymatic dissociation, organ on a chip, and bioprinting are being widely used to prepare organoids.
On the other hand, mechanical chopping provides metabolically active brain slices in various studies, but lacks the metabolic studies for organoid preparation. Our research has introduced an innovative method for generating patient-derived 3D organoids using an automated chopping machine. We demonstrated a nearly 70%reduction in manufacturing time and a significantly higher organoids count compared to the manual dissection.
Offering a more efficient method for drug and immunotherapy screening in glioblastoma patients. The advancement of our study significantly accelerates the organoid production process, and at the same time is able to maintain the main characteristics of the original tumors. And by reducing the time as well as the resources required for organoid generation, our approach enhances the feasibility of large scale drug and immunotherapy screening for glioblastoma patients.
To begin, transfer the tumor tissue immersed in 25 milliliters of hibernate A into a sterilized glass Petri dish under a laminar flow cabinet. Using a microscope carefully eliminate all the necrotic tissue, then use a scalpel and tissue forceps to dissect the blood vessels out. Cut the tumor tissue into one to two cubic centimeter pieces.
Transfer them into a plastic Petri dish containing three milliliters of H-GPSA medium, and place the dish on ice. To manually process the tumor, transfer the tumor piece to a fresh glass Petri dish. Using two scalpels, dissect the segment into 0.5 milliliter sections under a microscope.
To mechanically process the tumor, first, position the blade properly, then adjust the slice thickness to 0.45 to 0.50 millimeters, and fix the table release knob to start mode. Cut the Angros block into a two-centimeter long cylinder and glue it onto the circular plastic dish of the chopper. Using a scalpel, create a pit in the Angros cylinder and place the tumor tissue in the pit.
Then position the plastic disc onto the mounting disc of the cutting table, and press the reset button to start the chopping process. After the first round, rotate the mounting disc by 90 degrees and repeat again to cut the tissue into rectangular pieces. Remove the plastic disc, and then using a tissue spatula rotate only the tumor tissue by 90 degrees.
Return the plastic disc to the cutting table and press reset to continue cutting. Then using a five-milliliter pipette, transfer the processed material along with the medium to the petri dish. Place the dish immediately on ice.
To begin, obtain the tumor tissue from the patients and process it with a chopper. Tilt the Petri dish containing chopped tumor upwards to 45 degrees, and carefully remove 2.5 milliliters of the H-GPSA medium. To wash the tumor pieces, add two milliliters of RBC lysis buffer, and incubate in an orbital shaker at a slow speed.
Then, discard the lysis buffer completely. Next, add four milliliters of patient-derived organoid medium to the tissue pieces, and transfer the tissue chunks to an ultra low attachment six-well plate. Incubate the plate in an orbital shaker for two to four weeks.
Every two days, change half of the spent medium with a prewarmed fresh patient derived organoid medium. To prevent tissue hypoxia, transfer the patient derived organoids to a sterile glass petri dish and observe the tissue morphology under a microscope. Then using a scalpel, cut the adhesive tissue.
Patient derived organoids obtained using the chopper method, reached the desired rounded shape within one week as compared to the manually chopped ones. And the number of organoids formed was significantly higher using this method.
Overview
This study introduces an automated method to generate patient-derived glioblastoma 3-dimensional organoids using a tissue chopper. The method significantly reduces manufacturing time while maintaining the characteristics of the original tumor, enhancing drug and immunotherapy screening.
Key Study Components
Area of Science
- Neuroscience
- Oncology
- Organoid Technology
Background
- Patient-derived organoids are crucial for studying glioblastoma.
- Current methods include manual dissection and enzymatic dissociation.
- 3D organoids better replicate tumor complexity for drug testing.
- Mechanical chopping has been underutilized in organoid preparation.
Purpose of Study
- To improve the efficiency of generating patient-derived glioblastoma organoids.
- To reduce manufacturing time while preserving tumor characteristics.
- To facilitate effective drug and immunotherapy screening.
Methods Used
- Automated chopping machine for organoid generation.
- Manual and mechanical processing of tumor tissue.
- Incubation of organoids in patient-derived medium.
- Microscopic observation of tissue morphology.
Main Results
- Achieved a nearly 70% reduction in manufacturing time.
- Significantly higher organoid counts compared to manual methods.
- Maintained the main characteristics of original tumors.
- Enhanced feasibility for large-scale drug screening.
Conclusions
- The automated chopping method is efficient for organoid production.
- It supports the advancement of drug and immunotherapy testing.
- This approach could lead to better treatment strategies for glioblastoma patients.
Frequently Asked Questions
Executive Industry Relevance
Efficient generation of patient-derived glioma organoids is critical for translational oncology pipelines, enabling high-fidelity modeling of tumor heterogeneity and rapid preclinical screening. The automated tissue chopper method accelerates organoid production while preserving tumor characteristics, supporting scalable drug and immunotherapy evaluation. This innovation enhances predictive confidence and operational throughput at key discovery and lead identification inflection points.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Enables robust modeling of glioblastoma heterogeneity for target interrogation.
- Supports functional validation of therapeutic hypotheses in disease-relevant systems.
- Facilitates mechanistic de-risking by preserving original tumor features in organoids.
Screening & Assay Development
- Provides standardized, reproducible organoid batches for downstream compound screening.
- Delivers quantitative outputs on organoid yield and morphology for assay readiness.
- Reduces manual variability, enhancing scalability and platform reuse for screening campaigns.
Translational & Preclinical Research
- Aligns ex vivo models with patient tumor biology for translational biomarker exploration.
- Enables continuity from discovery through preclinical validation using patient-matched systems.
- Supports risk-adjusted advancement decisions by maintaining tumor-specific phenotypes.
Pipeline & Workflow Integration
This method integrates at the interface of early discovery and preclinical screening, bridging patient tissue acquisition to high-throughput drug testing workflows.
- Discovery Biology: Accelerates hypothesis testing and pathway clarification using patient-derived organoids.
- Screening: Delivers reproducible, quantitative organoid outputs for assay development and compound evaluation.
- Analytics: Enables statistical comparison of organoid yield, proliferation, and apoptosis across manufacturing methods.
- Translational Research: Maintains disease relevance for biomarker and therapeutic response studies.
- Enterprise Reuse: Establishes a scalable, standardized organoid generation capability for diverse R&D programs.
Operational & Enterprise Impact
- Scientific Value: Increases predictive confidence and reduces mechanistic ambiguity in glioma modeling.
- Operational Value: Achieves significant time savings and reproducibility in organoid manufacturing.
- Strategic Value: Improves go/no-go decision quality and capital efficiency in early-stage oncology pipelines.
- Portfolio Impact: Enables risk-adjusted prioritization and advancement of candidate therapies.
Implementation Considerations
- Requires expertise in tissue handling and organoid culture techniques.
- Needs access to automated tissue chopper instrumentation and analytical microscopy.
- Demands cross-team standardization for reproducible organoid preparation and assessment.
- Adaptation may be needed for different tumor types or tissue sources.
- Practical limitations include tissue availability and initial setup for automation.
