Tumor-LN-oC has successfully developed and optimized individual platform components and prepared them for their integration in the final prototype in the last months of the project.
Objective 1: We have designed and fabricated microfluidic chips with artificial cilia for continuous recirculatory flow for culturing tumor and LN cells and tissue samples. The chips were validated for long-term cell culture with cell lines and surgical samples.
Objective 2: Tumor-LN-oC consortium successfully integrated the EC-QCL laser source with the MIP spectroscopy module and completed comprehensive testing and calibration of the system. The integrated MIP system has demonstrated full functionality within the Tumor-LN-oC platform, including successful analysis of tissues and cells in fluidic environments.
Objective 3: We have isolated lymph-like media in the upper section of the Tumor-LN-oC chips; cytokines identified from the patient-derived samples were highly variable and demonstrated patient-specific expression. We have identified “druggable” targets for a more personalized approach in cancer treatment, with findings further implicating Jagged1 as a regulator of both the protein matrix surrounding tumors and cell movement in breast cancer.
Objective 4: The micro-optics module, based on microlens arrays, creates images with an FOV of 8.4 x 6 mm and an optical resolution of 3.9 μm. The high-level control software integrates all system functionalities. Machine learning was used for cell segmentation and motion quantification, enabling real-time monitoring of directional cell motion. Extensive user feedback has resulted in a software package which can be deployed for multiple days to acquire migration data from the microfluidic channels.
Objective 5: All modules were integrated into a functional TRL5 Tumor–LN-on-Chip prototype, with demonstrated troubleshooting, stable fluidic control and long term live cell culture across multiple chip configurations and cell/tissue types. The platform supported real-time imaging of single cell migration, maintained viable cultures under flow, and enabled mid-infrared photothermal spectroscopy in microfluidic chips.
Objective 6: Tumor-LN-oC platform underwent extensive optimization to support metastasis diagnosis and molecular characterization of migrating tumor cells. Cells cultured on the microfluidic platform remained significantly healthier than those in traditional Transwell (TW) systems. These results confirm that tumour slices cultured under continuous flow on the Tumor-LN-oC generate sufficient, high-quality mRNA for downstream analysis of migrating versus non migrating tumour cells.
Objective 7: The regulatory roadmap for metastasis diagnosis and drug testing applications was generated, identifying applicable guidelines and technical standards. 2 meeting with the EMA’s innovation task force were held. A comprehensive regulatory whitepaper summarizes lessons learned and outlining the standards, guidelines, and strategic pathways relevant to future market authorization. The exploitation and commercialization plan provides a foundation for future exploitation and continued cooperation of partners.
