Maintenance of steroid metabolism and hormone responsiveness in cryopreserved dog, monkey and human hepatocytes.
The efficient and effective use of hepatocytes from larger species and rare human material requires a reliable storage method for cells not needed on the day of preparation. Cryopreservation would seem to be the only viable alternative. In this study the suitability of a published cryopreservation technique on dog, monkey and human hepatocytes has been examined and the cells were tested for functionality directly after thawing and subsequent to culture using steroid metabolism and hormone responsiveness of glycogen phosphorylase a. Monkey and human hepatocytes appear to survive the freezing and thawing process better than dog cells-the latter losing the ability to respond to adrenergic stimuli and their ability to maintain steroid metabolism in culture. Although monkey and human cells do preserve their steroid metabolising capacity after freeze/thawing, there is not the significant increase in enzyme activity seen during culturing freshly isolated cells. It would appear, therefore, that some damage has occurred to the cells during the freeze/thaw process. As previously noted, Williams' medium E is superior to Ham's F-10 in maintaining enzyme activities in culture. It is suggested that cryopreservation is the way forward for the development of stockpiles of viable hepatocytes for biomedical and toxicological research and development but that further modifications to the process are still necessary to optimise the maintenance of liver-specific functions in the thawed cells.
Skett
P
P
Institute of Biomedical and Life Sciences, University of Glasgow, Scotland, UK. p.skett@bio.gla.ac.uk
Roberts
P
P
Khan
S
S
D000735:Androstenedione / Q000378:metabolism*; D000818:Animals; D002470:Cell Survival; D015925:Cryopreservation; D004285:Dogs; D004790:Enzyme Induction / Q000187:drug effects; D006801:Humans; D007545:Isoproterenol / Q000494:pharmacology; D008099:Liver / Q000378:metabolism; D008252:Macaca fascicularis; D008297:Male; D009926:Organ Preservation; D010656:Phenylephrine / Q000494:pharmacology; D006005:Phosphorylases / Q000378:metabolism*; D013045:Species Specificity
D010656:Phenylephrine; D000735:Androstenedione; D006005:Phosphorylases; D007545:Isoproterenol