Related Experiment Video
Updated: Jul 31, 2025
A High-Throughput Platform for Culture and 3D Imaging of Organoids
Published on: October 14, 2022
3D dynamic cultures of HGSOC organoids to model innovative and standard therapies
Enrico Cavarzerani1,2, Isabella Caligiuri1, Michele Bartoletti3
1Pathology Unit, Centro di Riferimento Oncologico di Aviano (C.R.O.) IRCCS, Aviano, Italy.
Abstract:
High-grade serous ovarian cancer (HGSOC) needs new technologies for improving cancer diagnosis and therapy. It is a fatal disease with few options for the patients. In this context, dynamic culture systems coupling with patient-derived cancer 3D microstructures could offer a new opportunity for exploring novel therapeutic approaches. In this study, we optimized a passive microfluidic platform with 3D cancer organoids, which allows a standardized approach among different patients, a minimum requirement of samples, multiple interrogations of biological events, and a rapid response. The passive flow was optimized to improve the growth of cancer organoids, avoiding the disruption of the extracellular matrix (ECM). Under optimized conditions of the OrganoFlow (tilting angle of 15° and an interval of rocking every 8 min), the cancer organoids grow faster than when they are in static conditions and the number of dead cells is reduced over time. To calculate the IC values of standard chemotherapeutic drugs (carboplatin, paclitaxel, and doxorubicin) and targeted drugs (ATRA), different approaches were utilized. Resazurin staining, ATP-based assay, and DAPI/PI colocalization assays were compared, and the IC values were calculated. The results showed that in the passive flow, the IC values are lower than in static conditions. FITC-labeled paclitaxel shows a better penetration of ECM under passive flow than in static conditions, and cancer organoids start to die after 48 h instead of 96 h, respectively. Cancer organoids are the last frontiers for ex vivo testing of drugs that replicate the response of patients in the clinic. For this study, organoids derived from ascites or tissues of patients with Ovarian Cancer have been used. In conclusion, it was possible to develop a protocol for organoid cultures in a passive microfluidic platform with a higher growth rate, faster drug response, and better penetration of drugs into ECM, maintaining the samples' vitals and collecting the data on the same plate for up to 16 drugs.
Insights
This study optimized a passive microfluidic platform for 3D ovarian cancer organoids, improving drug testing. The new method enhances organoid growth and drug penetration for faster, more accurate therapeutic evaluations.
Area of Science:
- Biotechnology
- Oncology
- Microfluidics
Background:
- High-grade serous ovarian cancer (HGSOC) is a fatal disease with limited therapeutic options.
- Dynamic culture systems with patient-derived 3D cancer microstructures offer novel therapeutic exploration.
- Existing methods require significant samples and lack rapid response for drug screening.
Purpose of the Study:
- To optimize a passive microfluidic platform for culturing 3D ovarian cancer organoids.
- To evaluate the platform's efficacy in improving organoid growth, drug penetration, and response time.
- To establish a standardized, rapid, and sample-efficient method for ex vivo drug testing in HGSOC.
Main Methods:
- Optimization of a passive microfluidic platform (OrganoFlow) with specific tilting angles and rocking intervals.
- Culturing of patient-derived ovarian cancer organoids under static and optimized passive flow conditions.
- Assessment of organoid growth, cell viability (Resazurin, ATP, DAPI/PI assays), and drug penetration (FITC-labeled paclitaxel).
Main Results:
- Optimized passive flow conditions (15° tilt, 8 min rocking interval) enhanced organoid growth rate and reduced cell death compared to static culture.
- Lower IC50 values for chemotherapeutic and targeted drugs were observed in passive flow conditions, indicating increased drug efficacy.
- Improved penetration of FITC-labeled paclitaxel into the extracellular matrix (ECM) and accelerated cancer cell death (48h vs. 96h) were noted under passive flow.
Conclusions:
- The developed passive microfluidic platform protocol supports enhanced organoid culture with faster growth and drug response.
- The platform facilitates better drug penetration into the ECM and provides a standardized approach for ex vivo drug screening.
- This technology enables rapid, sample-efficient evaluation of multiple drugs, potentially improving therapeutic strategies for ovarian cancer patients.
