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Cryopreservation of precision-cut rat liver slices using a computer-controlled freezer

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Author: Maas, W.J.M. · Leeman, W.R. · Groten, J.P. · Sandt, J.J.M. van de
Institution: Centraal Instituut voor Voedingsonderzoek TNO
Source:Toxicology in Vitro, 6, 14, 523-530
Identifier: 41720
doi: doi:10.1016/S0887-2333(00)00042-4
Keywords: Computer-controlled freezing · Cryopreservation · Rat liver slices · Animal tissue · Computer · Controlled study · Cryopreservation · Dehydration · Liver slice · Nonhuman · Rat · Species difference · Technique · Thawing · Toxicity testing · Adenosine Triphosphate · Animal · Cell Separation · Cell Survival · Computer Systems · Cryopreservation · Dinitrochlorobenzene · Formazans · Freezing · Glutathione · Glutathione Transferase · Hepatocytes · Lactate Dehydrogenase · Liver · Male · Organ Preservation · Proteins · Rats · Rats, Wistar · Support, Non-U.S. Gov't · Testosterone · Tetrazolium Salts · Time Factors · Urea


Precision-cut liver slices are frequently used to study hepatic toxicity and metabolism of xenobiotics in vitro. Successful cryopreservation techniques will enhance an efficient and economic use of scarcely available (human) liver tissue. For primary hepatocytes, slow freezing has been accepted as the best approach towards successful cryopreservation. For slices, however, no agreement exists on the optimal way of cryopreservation and both slow and fast freezing techniques have been reported. The aim of the present study was to determine the applicability of a computer-controlled slow freezing technique for the cryopreservation of (rat) liver slices. Thus far, this technique has not been described in detail. Our studies confirmed that slow freezing was most successful in the cryopreservation of primary rat hepatocytes. Based on this observation, the slow freezing technique was applied to the cryopreservation of rat liver slices. Directly after thawing, slice viability was between 60 and 100% of fresh values, depending on the parameter determined. However, after additional culturing, slice viability was reduced. This decrease in slice viability was more pronounced in comparison to primary hepatocytes. In conclusion, the slow freezing technique was confirmed to be a successful approach for the cryopreservation of primary rat hepatocytes, and was found to be of limited use for the cryopreservation of rat liver slices. Copyright (C) 2000 Elsevier Science Ltd. Chemicals/CAS: Adenosine Triphosphate, 56-65-5; Dinitrochlorobenzene, 97-00-7; Formazans; Glutathione Transferase, EC; Glutathione, 70-18-8; Lactate Dehydrogenase, EC; MTT formazan, 23305-68-2; Proteins; Testosterone, 57-85-2; Tetrazolium Salts; Urea, 57-13-6