1: Cell Physiol Biochem.  2003;13(2):103-12.  

Cryopreservation of rat precision-cut liver slices is associated with major
metabolic stress and ionic perturbations.

Vanhulle VP, Martiat GA, Bontemps F, Vincent MF, Pycke JM, Verbeeck RK, Horsmans
Y, Delzenne N.

Unit of Pharmacokinetics, Metabolism, Nutrition and Toxicology, Universite
Catholique de Louvain, Belgium.

BACKGROUND/AIMS: Cryopreserved precision-cut liver slices (PCLS) in culture
exhibit a rapid decline in ATP level and anabolic processes by unknown
mechanisms. The aim of this study is to elucidate the key events explaining the
alterations occurring in cryopreserved PCLS. METHODS: Glucose metabolism,
mitochondrial activities, ionic homeostasis and caspase-3 activity were assessed
in fresh or cryopreserved (rapid freezing-thawing conditions) rat PCLS within
the first hour of incubation in Williams' medium E at 37 degrees C. RESULTS:
Despite a similar glycolytic activity under both conditions, only fresh PCLS
were able to gradually recover their ATP and potassium content. Glycogen content
dropped more rapidly in cryopreserved than in control PCLS. Only cryopreserved
PCLS exhibited a decline in O2 consumption and a lower ATP/ADP ratio from 30 min
of incubation with a loss of coupling of oxygen consumption to ATP synthesis and
mitochondrial calcium transport after 1 hour of incubation. Caspase-3 activation
was already present in cryopreserved PCLS after a few minutes of incubation.
CONCLUSIONS: The lack of restoration of potassium/sodium exchange might be a
primary event in the metabolic alterations occurring in cryopreserved PCLS after
thawing, that contributes to further mitochondrial alterations, caspase
activation and compromises ATP-dependent anabolic pathways.

PMID: 12649595 [PubMed - indexed for MEDLINE]