X-Message-Number: 29517 From: David Stodolsky <> Subject: Cryopreservation of Primary Porcine Hepatocytes Date: Sun, 20 May 2007 21:28:14 +0200 --Apple-Mail-8--481368245 delsp=yes; format=flowed PhD Defendant: Cand. med. vet. Anne-Dorthe Bonde Loven Time and Place: Friday 25 May 2007 at 1:30 p.m. in Lecture Hall 3-14, Thorvaldsenvej 40, C. Subject Area: Toxicology Title of PhD Thesis: Cryopreservation of Primary Porcine Hepatocytes Title of Lecture: Cryopreservation of Primary Porcine Hepatocytes Summary: The aim of this thesis was to compare the metabolic capacity and the carbohydrate metabolism in cyopreservated porcine hepatocytes with those obtained in freshly isolated cells in order to examine cryopreservated cells as a tool in the drug screening procedures. Cryopreservation caused a decrease in both phase I and II metabolism. Thus, CYP2A activity in terms of hydroxylation of coumarin to 7-hydroxy coumarin decreased two-thirds compared to freshly isolated cells, whereas CYP3A activity in terms of 6 -hydroxylation of testosterone and UGT activity in terms of glucuronidation of p-nitrophenol both decreased to about half the values in fresh cells. Carbohydrate metabolism was also reduced by cryopreservation: glycogen synthesis was decreased to less than half and glucose produced from glyco- genolysis and gluconeogenesis was decreased with 25-50%. Other effects of cryopreservation included tendencies to decreased attachment and viability. Co-culturing with epithelial cells did not improve the function of cryopreserved cells.In conclusion cryopreserved primary porcine hepatocytes are useful in culture but have a general decrease of activity compared to similar fresh hepatocytes. Principal Supervisor: Professor Christian Friis, Toxicology, Department of Veterinary Pathology, Faculty of Life Sciences, University of Copenhagen. Co-Supervisor: Principal Scientist, Anette Kristensen, Novo Nordisk Research Scientist, Birgitte Andersen, Novo Nordisk Examination Committee: European Scientific Consultant Nicola Hewitt, CellzDirect. Associated Professor Dr. phil Niels Grunnet, Panum Institute, Associated Professor Mette Tinglev Skaanild, Department of Veterinary Pathology, Faculty of Life Sciences, University of Copenhagen. --Apple-Mail-8--481368245 Content-Transfer-Encoding: base64 [ AUTOMATICALLY SKIPPING BASE64 ENCODING! ] Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=29517