MSc candidate University of Toronto Toronto, Canada
Abstract: In our laboratory, the primary objective is to engineer a transplantable human organ using a patient's stem cells and decellularized pig organs as a scaffold, with the goal of addressing the growing demand for organ transplants. A critical aspect of this work is normothermic ex-vivo perfusion (NEVP) culture, which supports organ growth and maintenance. The challenge lies in the high cost and complexity of current clinical NEVP systems, which rely heavily on external oxygen tanks, red blood cells, and expensive single-use components. Once the NEVP system is fully developed and optimized, the next phase will focus on humanizing pig kidneys with patient-derived induced pluripotent stem cells (iPSCs), advancing us toward functional, transplantable organs.
To address these issues, we have designed an affordable and efficient normothermic ex-vivo perfusion (NEVP) system using a native pig kidney model. This system is based on a mouse NEVP system developed in our lab. Key innovations include the elimination of an external oxygen source and red blood cells, reusable accessories, and optimized medium flow. All of these leads to reducing costs and enhancing accessibility. Additionally, we implemented contamination prevention strategies and fine-tuned osmolarity to reduce edema and therefore prevent cell death. Pressure, flow, and oxygen sensors were incorporated to ensure physiologically relevant conditions .
Our results demonstrate successful maintenance of a native pig kidney for 24 hours using this new system, with kidney health assessed by urine analysis, including total protein, albumin and creatinine levels in the ultrapurfusate (urine). Moving forward, we plan to optimize oncotic pressure and investigate the use of the oxygen carrier perfluorocarbon (PFC) to extend the kidney’s viability. This work represents a significant step toward making ex-vivo perfusion more cost-effective and scalable for both organ transplant applications and research studies that require organ culture.
