Document Detail


Increased post-thaw viability and phase I and II biotransformation activity in cryopreserved rat liver slices after improvement of a fast-freezing method.
MedLine Citation:
PMID:  10950856     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
An existing cryopreservation method for liver slices applies 12% dimethylsulfoxide and rapid freezing. We found that cells in rat liver slices cryopreserved in this manner deteriorated rapidly upon culturing. To improve this cryopreservation method, we varied the dimethylsulfoxide concentration (0, 12, 18, and 30%), the cryopreservation medium (Williams medium E, fetal calf serum, and University of Wisconsin medium), slice thickness, and the storage period at 4 degrees C during slice preparation before cryopreservation. After thawing, slices were cultured for 4 h at 37 degrees C before their viability was evaluated by their potassium content and the number of intact cells determined histomorphologically. The biotransformation capacity of liver slices cryopreserved by the improved method was assessed by testosterone oxidation, hydroxycoumarin sulfation, and glucuronidation. Best results were obtained with 18% dimethylsulfoxide in Williams medium E: the potassium content of cryopreserved slices was higher than 65%, and the number of intact cells was higher than 60% of that in fresh slices; with 12% dimethylsulfoxide, potassium content was less than 40%, and the number of intact cells was less than 30%. Results did not differ between the three cryopreservation media. Viability of thin slices (8-10 cell layers) was better maintained than that of thicker slices (>14 cell layers). Storage at 4 degrees C of slices before cryopreservation decreased viability after cryopreservation. Both oxidative and conjugation activities were better than 60% of fresh values. Although results varied, slices cryopreserved with this improved method and cultured for 4 h retained viability between 50 and 80%, and biotransformation activity between 60 and 90% of fresh slices.
Authors:
I A de Graaf; D van der Voort; J H Brits; H J Koster
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Drug metabolism and disposition: the biological fate of chemicals     Volume:  28     ISSN:  0090-9556     ISO Abbreviation:  Drug Metab. Dispos.     Publication Date:  2000 Sep 
Date Detail:
Created Date:  2000-10-04     Completed Date:  2000-10-04     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  9421550     Medline TA:  Drug Metab Dispos     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1100-6     Citation Subset:  IM    
Affiliation:
Solvay Pharmaceuticals BV, Drug Safety Department, The Netherlands. inge.degraaf@solvay.com
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MeSH Terms
Descriptor/Qualifier:
Animals
Biotransformation
Cell Survival / drug effects
Chromatography, High Pressure Liquid
Cold Temperature
Cryopreservation / methods,  standards*
Dimethyl Sulfoxide / pharmacology
Freezing*
Liver / cytology,  drug effects,  metabolism*
Male
Potassium / metabolism
Rats
Rats, Wistar
Temperature
Testosterone / pharmacokinetics
Time Factors
Tissue Preservation
Chemical
Reg. No./Substance:
58-22-0/Testosterone; 67-68-5/Dimethyl Sulfoxide; 7440-09-7/Potassium

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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