Cryopreservation of precision-cut rat liver slices using a computer-controlled freezer.
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.
Maas
W J
WJ
TNO Nutrition and Food Research, Department of Explanatory Toxicology, PO Box 360, 3700, AJ Zeist, The Netherlands. maas@voeding.tno.nl
Leeman
W R
WR
Groten
J P
JP
van de Sandt
J J
JJ
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002469:Cell Separation; D002470:Cell Survival; D003199:Computer Systems; D015925:Cryopreservation / Q000295:instrumentation* / Q000379:methods; D004137:Dinitrochlorobenzene / Q000378:metabolism; D005562:Formazans / Q000378:metabolism; D005615:Freezing; D005978:Glutathione / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000378:metabolism; D008297:Male; D009926:Organ Preservation / Q000295:instrumentation* / Q000379:methods; D011506:Proteins / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013739:Testosterone / Q000378:metabolism; D013778:Tetrazolium Salts / Q000378:metabolism; D013997:Time Factors; D014508:Urea / Q000378:metabolism
D004137:Dinitrochlorobenzene; D005562:Formazans; D011506:Proteins; D013778:Tetrazolium Salts; C079782:MTT formazan; D013739:Testosterone; D000255:Adenosine Triphosphate; D014508:Urea; D007770:L-Lactate Dehydrogenase; D005982:Glutathione Transferase; D005978:Glutathione