Macrosteatotic hepatocyte defatting

Since there is a shortage of transplantable livers, the donor pool needs to be increased. One method to accomplish this is to recondition extended criteria donor grafts, a large portion of which are moderate to severe macrosteatotic livers. Transplantation of these livers often leads to primary nonfunction caused by an increased susceptibility to the effects of ischemia reperfusion injury (I/R) that result from the harvesting, transportation, and transplantation of the liver. Our lab has developed a novel procedure to recondition these livers through an ex vivo perfusion that will reduce the hepatic triglyceride content before the onset of the effects of I/R injury. To be feasibly performed in a clinical setting, the defatting must be completed in a matter of hours. While attempts to identify the ideal defatting cocktail in static culture only resulted in defatting livers after days of treatment, the rate of defatting increases to more a clinically relevant timeframe in a flow environment. My project focuses on understanding the differences between a static and flow environment and using this information to develop the ideal parameters for defatting. I hypothesize that using flow and appropriate defatting agents, steatotic hepatocytes can be defatted in a clinically relevant time of hours, without harming viability or function, to a level that reduces or eliminates increased sensitivity to I/R injury. In order to prove the hypothesis I will use the following 3 specific aims: 1. Develop a metabolic flux analysis model to explore defatting metabolic pathways. 2. Develop a computational fluid dynamics and kinetic model of a vitro flow reactor to optimize key transport parameters. 3. Validate the optimal conditions of flow has on the viability and function in hepatocyte cell lines.