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Hypothermic preservation of hepatocytes on gelatin gels
Authors:Peter J. Evans
Affiliation:1. AGH University of Science and Technology, Department of Environmental Management and Protection, al. Mickiewicza 30, 30-059 Kraków, Poland;2. University of Agriculture in Kraków, Department of Forest Ecology and Reclamation, Institute of Ecology and Silviculture, Faculty of Forestry, al. 29 Listopada, 31 – 425 Kraków, Poland
Abstract:Hypothermic preservation of hepatocytes on gelatin gels allows hepatocytes to be stored for at least 9 days. The procedure is easy, inexpensive and does not require specialised equipment. The cells retain their morphology and are released as separate spherical entities by dissolving the gelatin layer at 37°. The recovered cells have an intact plasma membrane as judged by lactate dehydrogenase activity, attachment efficiency and subsequent monolayer formation in culture. Functional tests show that the cells recover quickly from the storage conditions. Rates of protein synthesis are maintained over a 6 day period and remain at 62% of the initial level on day 9. The liver specific, hormonal induction of the enzyme tyrosine aminotransferase is apparent throughout the culture period. In addition a phenotypic marker enzyme, γ-glutamyl transpeptidase, remained at basal levels.
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