Cold-preservation of primary isolated pig hepatocytes – Fundamental study concerning optimal medium – *
As a source for liver-targeted regeneration medicine, a supply of normal human hepatocytes is essential. However there are limitations to the supply of human liver from surgical resection. Therefore, the efficient use of a limited liver supply is required. For this purpose it is extremely important to establish efficient techniques for the cell isolation and cold-preservation of isolated liver cells. As for the isolation of hepatocytes from surgically removed pig livers, dispase perfusion followed by collagenase digestion should yield hepatocytes with more than 90% viability. Cold-preservation for eight hours using isolated pig hepatocytes in the following preservation media was compared ; 1) University of Wisconsin (UW) solution with ascorbic acid 2 glucocide (AA 2 G) 2) University of Wisconsin (UW) solution 3) 100% fetal bovine serum 4) Dulbecco's Modified Eagle's Medium supplemented with 10% fetal bovine serum With the UW solution, viability, plating efficiency and ammonium clearance were well preserved after eight hours of cold-preservation. UW +AA 2 G solution yielded the best preservation effect. The use of AA 2 G inhibited activation of caspase-3, resulting in well-preserved cellular ATP levels. Based on the above results, the UW +AA 2 G solution appeared to be most suitable for cold-preservation of pig hepatocytes. Clininal application for the cold-preservation of human hepatocytes can be expected. (Accepted on Augusut 28, 2002) Kawasaki Igakkaishi 28(3) : 199-208, 2002
