Primary Culture of Rat Hepatocytes After Cold Storage in the University of Wisconsin Solution: A Tool to Study the Effects of Hypothermic Preservation

Abstract

The effect of long-term hypothermic preservation on the function and morphology of rat hepatocytes cultured up to 120 h was studied. Cold storage was performed using modified University of Wisconsin solution, originally developed for organ preservation. After different times of cold storage of hepatocytes, a primary culture system was used to evaluate their ability to attach and form a monolayer of cells on a Petri dish, as well as to perform metabolic functions: capacity of maintaining intracellular enzymes, ammonium removal, and presence of albumin expression. The primary culture system is good enough to evaluate the effects on cells maintained in hypothermia when these cells are placed in normothermic conditions in an oxygenated medium. The results showed that preserved hepatocytes maintained relatively good functions (expressed as attachment, ammonia removal, and albumin expression) when they were preserved up to 72 h in modified University of Wisconsin solution at 0°C and then cultured for up to 120 h. This work raises the possibility of efficiently studying the functional response of cold-stored cells and provides the potential of having viable and functional cells to be applied in different systems such as bioartificial liver devices or cellular transplantation, overcoming, in part, the scarcity of liver donors.