PMID- 11350010
OWN - NLM
STAT- MEDLINE
DCOM- 20010524
LR  - 20190701
IS  - 0024-3205 (Print)
IS  - 0024-3205 (Linking)
VI  - 68
IP  - 21
DP  - 2001 Apr 13
TI  - Cryopreservation of rat precision-cut liver slices by ultrarapid freezing: 
      influence on phase I and II metabolism and on cell viability upon incubation for 
      24 hours.
PG  - 2391-403
AB  - Several cryopreservation methods for precision-cut rat liver slices (PCLS) have 
      been proposed, allowing a short-term (a few hours) maintainance of viability and 
      functionality upon thawing. The aim of the present study was to test the 
      metabolic capacity of PCLS cryopreserved by an ultrarapid method. The 
      biotransformation of paracetamol to its glucuronide and sulfate conjugates and of 
      midazolam to its hydroxylated metabolites was studied in thawed PCLS incubated 
      for 24 hours at 37 degrees C in Williams' medium E. In addition, protein levels 
      of the key enzymes involved in these metabolic reactions, i.e. UGT1A1, ST1A1, 
      CYP2E1 and CYP3A2 were determinated. In addition, biological markers of cell 
      function (ATP and glycogen levels) and toxicity (LDH leakage in the medium) were 
      also measured. Compared to controls (non cryopreserved PCLS), CYP3A2 activity and 
      content and CYP2E1 content were maintained at the same level all along the 
      incubation, whereas paracetamol glucuronidation and sulfation dropped to 24 and 
      21% of the control value, respectively, immediately after thawing. 
      Freezing-thawing conditions also modified cell functionality, leading to a lower 
      intracellular ATP and glycogen content, and an increase in cell lysis, as shown 
      by LDH released in the medium. The results of this study suggest that 
      cryopreserved PCLS are able to maintain some phase I activities for 24 hours 
      after thawing whereas some phase II metabolic capacities are not maintained.
FAU - Vanhulle, V P
AU  - Vanhulle VP
AD  - Unit of Pharmacokinetics, Metabolism, Nutrition and Toxicology, Universite 
      Catholique de Louvain, Brussels, Belgium.
FAU - Martiat, G A
AU  - Martiat GA
FAU - Verbeeck, R K
AU  - Verbeeck RK
FAU - Horsmans, Y
AU  - Horsmans Y
FAU - Calderon, P B
AU  - Calderon PB
FAU - Eeckhoudt, S L
AU  - Eeckhoudt SL
FAU - Taper, H S
AU  - Taper HS
FAU - Delzenne, N
AU  - Delzenne N
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - Life Sci
JT  - Life sciences
JID - 0375521
RN  - 0 (Isoenzymes)
RN  - 362O9ITL9D (Acetaminophen)
RN  - 8L70Q75FXE (Adenosine Triphosphate)
RN  - 9005-79-2 (Glycogen)
RN  - 9035-51-2 (Cytochrome P-450 Enzyme System)
RN  - EC 1.1.1.27 (L-Lactate Dehydrogenase)
RN  - R60L0SM5BC (Midazolam)
SB  - IM
MH  - Acetaminophen/pharmacokinetics
MH  - Adenosine Triphosphate/metabolism
MH  - Animals
MH  - Biotransformation
MH  - Blotting, Western
MH  - Cell Survival/drug effects/physiology
MH  - Cryopreservation/*methods
MH  - Cytochrome P-450 Enzyme System/metabolism
MH  - *Freezing
MH  - Glycogen/metabolism
MH  - Hepatocytes/cytology/drug effects/metabolism
MH  - Isoenzymes/metabolism
MH  - L-Lactate Dehydrogenase/metabolism
MH  - *Liver
MH  - Male
MH  - Midazolam/pharmacokinetics
MH  - Organ Culture Techniques
MH  - Organ Preservation/*methods
MH  - Rats
MH  - Rats, Wistar
EDAT- 2001/05/15 10:00
MHDA- 2001/05/26 10:01
CRDT- 2001/05/15 10:00
PHST- 2001/05/15 10:00 [pubmed]
PHST- 2001/05/26 10:01 [medline]
PHST- 2001/05/15 10:00 [entrez]
AID - S0024320501010311 [pii]
AID - 10.1016/s0024-3205(01)01031-1 [doi]
PST - ppublish
SO  - Life Sci. 2001 Apr 13;68(21):2391-403. doi: 10.1016/s0024-3205(01)01031-1.