The effect of rewarming media composition on the ammonia detoxification ability of cold preserved rat hepatocytes

Abstract

We examined how different media composition of rewarming solutions affected ammonium detoxification function, urea synthesis and the viability of hepatocytes after 72 hs of cold storage in UW solution. Freshly isolated rat hepatocytes were incubated at 37 ºC in a cell culture medium (MEM-E) with 3 mM glycine, 5 mM fructose and 2.5 mM adenosine (group 1) and in Krebs-Heinseleit buffer with 3 mM glycine, 5 mM fructose, 2 mM ornithine, 10 mM lactate and adenosine, that was used in two different concentrations: 2.5 mM (group 2) and 10 mM (group 3). We found that freshly isolated cells produced ammonium in group 1 and 2 but the cells were able to diminish ammonium extracellular concentration in group 3. Urea synthesis and ammonium extracellular concentration in group 1 was higher than in group 2. As a result of this observations, we used the Krebs-Heinseleit solution with addition of 10 mM adenosine to determinate the effect of hypothermic preservation on ammonium detoxification and urea synthesis ability of cells. In conclusion the addition of 2.5 mM adenosine into the rewarming medium interfered with the detection of ammonium detoxification of hepatic cells.