Related Experiment Video
Updated: Jul 26, 2025
Multiparametric Tumor Organoid Drug Screening Using Widefield Live-Cell Imaging for Bulk and Single-Organoid Analysis
Published on: December 23, 2022
A perfused multi-well bioreactor platform to assess tumor organoid response to a chemotherapeutic gradient
Elisa Marie Wasson1, Wei He2, Jesse Ahlquist1
1Materials Engineering Division, Lawrence Livermore National Laboratory, Livermore, CA, United States.
Abstract:
There is an urgent need to develop new therapies for colorectal cancer that has metastasized to the liver and, more fundamentally, to develop improved preclinical platforms of colorectal cancer liver metastases (CRCLM) to screen therapies for efficacy. To this end, we developed a multi-well perfusable bioreactor capable of monitoring CRCLM patient-derived organoid response to a chemotherapeutic gradient. CRCLM patient-derived organoids were cultured in the multi-well bioreactor for 7 days and the subsequently established gradient in 5-fluorouracil (5-FU) concentration resulted in a lower IC50 in the region near the perfusion channel versus the region far from the channel. We compared behaviour of organoids in this platform to two commonly used PDO culture models: organoids in media and organoids in a static (no perfusion) hydrogel. The bioreactor IC50 values were significantly higher than IC50 values for organoids cultured in media whereas only the IC50 for organoids far from the channel were significantly different than organoids cultured in the static hydrogel condition. Using finite element simulations, we showed that the total dose delivered, calculated using area under the curve (AUC) was similar between platforms, however normalized viability was lower for the organoid in media condition than in the static gel and bioreactor. Our results highlight the utility of our multi-well bioreactor for studying organoid response to chemical gradients and demonstrate that comparing drug response across these different platforms is nontrivial.
Insights
Developing new colorectal cancer liver metastases (CRCLM) therapies requires better preclinical models. A novel perfusable bioreactor effectively monitors CRCLM patient-derived organoid responses to chemotherapy gradients, aiding drug screening.
Area of Science:
- Oncology
- Biomedical Engineering
- Drug Discovery
Background:
- Colorectal cancer liver metastases (CRCLM) present a significant therapeutic challenge, necessitating advanced preclinical models for drug development.
- Current patient-derived organoid (PDO) culture methods may not accurately reflect in vivo drug responses due to lack of physiological conditions.
- There is a critical need for innovative platforms to assess therapeutic efficacy against CRCLM.
Purpose of the Study:
- To develop and validate a multi-well perfusable bioreactor system for evaluating CRCLM patient-derived organoids.
- To investigate the response of CRCLM organoids to a chemotherapeutic gradient within the bioreactor.
- To compare the drug response of organoids in the bioreactor with traditional culture methods (in media and static hydrogel).
Main Methods:
- Development of a multi-well perfusable bioreactor for culturing CRCLM patient-derived organoids.
- Establishment of a 5-fluorouracil (5-FU) concentration gradient within the bioreactor.
- Culturing organoids for 7 days and assessing their response to the 5-FU gradient.
- Comparison of IC50 values and normalized viability across bioreactor, static hydrogel, and media-only conditions.
- Utilizing finite element simulations to analyze drug dose delivery (AUC).
Main Results:
- The perfusable bioreactor demonstrated a gradient in 5-FU IC50 values, with lower values near the perfusion channel.
- Bioreactor IC50 values were significantly higher than those for organoids cultured in media alone.
- Organoids cultured in media showed lower normalized viability compared to those in the static gel and bioreactor.
- Finite element simulations indicated similar total drug dose (AUC) across platforms, but differential viability responses.
Conclusions:
- The developed multi-well perfusable bioreactor is a valuable tool for studying organoid responses to chemical gradients in CRCLM.
- Drug response comparisons across different preclinical platforms (bioreactor, static hydrogel, media) are complex and require careful consideration of experimental conditions.
- This platform offers a more physiologically relevant model for screening potential therapies for colorectal cancer liver metastases.
