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Cryopreserved hepatocyte monolayers

Stevenson, D. and Morgan, C. and Goldie, E.I. and Connel, G. and Grant, M.H. (2002) Cryopreserved hepatocyte monolayers. Biochemical Society Transactions, 30. p. 94. ISSN 0300-5127

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Abstract

Successful cryopreservation of hepatocytes would be useful to the pharmaceutical industry and for bioartificial liver support systems. Unfortunately, cryopreservation of hepatocytes in suspension results in low attachment efficiency in culture upon thawing. To circumvent this problem, we have frozen rat hepatocytes as monolayers on collagen substrates. Hepatocytes prepared from male Sprague Dawley rats by collagenase perfusion were cultured (3*106 cells/60mm Petri dish) on collagen films (30mg/cm2) in 2ml Chee's medium containing 5% v/v foetal calf serum (FCS). 24h cultures were frozen in Chee's medium with 10% Dimethyl Sulphoxide and between 0-90% FCS at -708C for 24h.