Project Summary/Abstract: TR&D-4 will demonstrate process automation on ARMI|BioFabUSA’s Tissue Foundry, a scalable, modular, automated and closed tissue manufacturing platform. The Tissue Foundry is the integration of primarily off-the- shelf components to perform the sequential, modular steps of producing a tissue. The Tissue Foundry automates thawing a vial of adherent cells, expanding them on microcarriers in stirred tank vessels, harvesting cells and exchanging cells into media promoting subsequent differentiation or tissue development steps, culturing the tissues during maturation, and preservation and packaging of the final product. To re-design the manual process to the closed, automated Tissue Foundry process, ARMI’s Deep Tissue Characterization Center deconstructs a manual process and develops tissue-type specific setpoints at both the endpoint and at intermediate stages of production using experimentation and modeling after deep multiparameter characterization of the cells and tissue. Once these setpoints are identified significant gaps remain in available tissue specific sensors and equipment to control and monitor tissue quality. Advanced sensor technologies (developed in TR&D-1 and –2) and prototype, actuator-enabled bioreactors (TR&D-3) will be integrated into the Tissue Foundry and demonstrated through production of CP target tissues in a tissue manufacturing relevant environment. This objective will be carried out in 3 specific aims: Aim 1: Physical integration of sensors with Tissue Foundry cell culture system. We will integrate newly-developed biosensors developed in TR&D-1, -2, and -3 into the Tissue Foundry for both measurement of parameters specific to the tissue process, and as a proof of concept for the feasibility of controlling these processes through feedback loops. Aim 2: Efficient control by integration of sensors and actuators with Tissue Foundry backbone automation and data management system. Tissue Foundry control is based on a supervisory control and data acquisition (SCADA) architecture and on the use of programmable logic controllers (PLCs), which will interface directly with sensors and actuators executing processing steps and real-time feedback control. Aim 3: Automated production of up to 10 tissues simultaneously. A manual tissue engineering process, starting with CCMEEC’s articular cartilage, will be assessed to identify optimal automation and sensor implementation. Once configured, the Tissue Foundry will be used to produce up to 10 tissues simultaneously using integrated bio-instructive bioreactors. Data from each step and the final product will be collected through integrated sensors, analysis of media aliquots, and destructive sampling to compare the manual and automated processes. TR&D4 will deliver an automated manufacturing system, leveraging the existing modules of the prototype Tissue Foundry, with vastly improved capabilities developed by TR&D-1, -2, and -3, for reliably producing complex tissues