Programmable Microfluidic Perfusion Bioreactor for 3D Organoid Culture and Screening Applications

Executive Summary of Predicate SBIR Phase I Grant and Team The overall objective of our predicate SBIR Phase I grant is to develop and validate a versatile, programmable microfluidic perfusion bioreactor platform for 3D organoid culture and screening applications. Nine out of ten drug candidates that demonstrate safety and efficacy in animal testing ultimately fail in human trials. Organoids offer a more representative and cost-effective alternative to traditional in vitro and animal models for drug screening, with the FDA Modernization Act 2.0 enabling this data to be used in drug approvals. As such, there is currently a large and growing unmet need for new bioreactor tools designed specifically for organoid culture. Mosaic Engineering is addressing this need by developing the Microlab, a low-cost perfusion bioreactor that leverages recent advances in MEMS technology to precisely drive fluids across a standardized microfluidic consumable interface. Our microfluidic consumables follow a standard SBS plate form factor, allowing integration into broader laboratory automation workflows. The specific aims of the Phase I project are to: (1) build a control manifold and automation software for microfluidic perfusion cell culture, (2) demonstrate 3D organoid differentiation with the perfusion culture system, and (3) demonstrate pharmacological dosing effects on cardiac organoids under perfusion. In partnership with Bullseye Biotechnologies, we will validate this platform by demonstrating drug testing with iPSC-derived cardiovascular organoids (CVOs) under perfusion culture. We have already developed an automated production process for manufacturing Bullseye’s differentiation kit and ECM patterned plate (A). We have also designed and produced a microfluidic chip with 6 organoid perfusion culture chambers with 5 addressable perfusion inlets (B). This chip attaches to a SBS-style reagent plate which attaches to a manifold and precision pressure controller (C-D). This system is compatible with conventional inverted microscopes (E-F). The Bullseye patterning method produces consistent organoids (G) which can be loaded into the perfusion chip and precisely dosed with drug candidates (H). The red outline in (B) indicates ROI shown in (H). The proposed i-Corps team includes: Frank Myers, PhD CEO & PI/PD Debkishore Mitra, PhD Technical Lead Oscar Abilez, MD, PhD Industry Expert All three team members are fully committed to meeting the time-intensive requirements of the training program (minimum 25 hours per week per team member) and attending all scheduled program events and webinars.