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Cryopreservation of hepatocyte monolayers

Stevenson, D.J. and Morgan, C. and Goldie, E.I. and Connel, G. and Grant, M.H. (2003) Cryopreservation of hepatocyte monolayers. In: Hepatocyte Users Group Meeting, 2003-03-28 - 2003-03-29, Aberdeen.

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Abstract

The ability to cryopreserve hepatocytes would be useful both to the pharmaceutical industry and for bioartificial liver support systems. Unfortunately, suspension cryopreservation protocols typically result in low attachment efficiencies of cells upon thawing. To circumvent this problem, we have frozen rat hepatocytes as monolayers on collagen substrates, and attempted to optimise this cryopreservation protocol. A variety of parameters were measured in non-frozen and post-thaw frozen monolayer cultures, including viability, total protein and intracellular reduced glutathione (GSH) concentration, kaempherol glucuronidation, and testosterone hydroxylation. The effect of altering cryopreservation media composition (% of foetal calf serum varying between 0-90%) or freezing (0.4-3.8ºC/min) and thawing rates (26-128ºC/min) on these parameters was investigated. Under optimal conditions, post thaw cryopreserved cells maintained 72±4% viability, 65±4% total protein, 46±8% GSH, 48±8% kaempherol glucuronidation, and 16±11% testosterone hydroxylation of their corresponding non-frozen controls (mean ±SEM, n=3). Cryopreservation of hepatocyte monolayers as opposed to suspensions results in a more representative population of cells, with high viability, function, and recovery rates.

Item type: Conference or Workshop Item (Paper)
ID code: 8435
Keywords: cryopreservation, monolayer hepatocyte cultures, carboxyfluorescein diacetate, cytochrome P-450 dependent monooxygenase activity, glucuronidation, Bioengineering
Subjects: Technology > Engineering (General). Civil engineering (General) > Bioengineering
Department: Faculty of Engineering > Bioengineering
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Depositing user: Strathprints Administrator
Date Deposited: 11 Aug 2009 11:54
Last modified: 17 Jul 2013 15:14
URI: http://strathprints.strath.ac.uk/id/eprint/8435

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