1: Life Sci. 2006 Feb 28;78(14):1570-7. Epub 2005 Oct 19. 

Role of apoptotic signaling pathway in metabolic disturbances occurring in liver
tissue after cryopreservation: Study on rat precision-cut liver slices.

Vanhulle VP, Neyrinck AM, Pycke JM, Horsmans Y, Delzenne NM.

Unit of Pharmacokinetics, Metabolism, Nutrition and Toxicology, Universite
Catholique de Louvain, 7369, Avenue E. Mounier 73, B-1200 Brussels, Belgium.

Precision-cut liver slices in culture (PCLS) appears as a useful and widely used
model for metabolic studies; the interest to develop an adequate
cryopreservation procedure, which would allow maintaining cell integrity upon
incubation, is needed to extend its use for human tissues. We have previously
shown that cryopreservation of rat PCLS leads to caspase-3 activation and early
alterations of their K(+) content upon incubation. In this study, we tested the
hypothesis that counteracting intracellular K(+) loss and/or interfering with
cell death signaling pathways could improve the viability of cryopreserved PCLS.
PCLS were prepared from male Wistar rat liver and cryopreserved by rapid
freezing before incubation. The addition of a caspase inhibitor-Z-DEVD-FMK (2.5
muM)-in the culture medium did not improve viability of cryopreserved PCLS.
Incubation of cryopreserved PCLS in a K(+) rich medium (135 mM) increased K(+)
content and avoided caspase-3 activation, but did not improve cell viability.
Caspase-3 inhibition, a decrease in cell lysis, and improvement of glycogen
content were observed in cryopreserved PCLS after addition of LiCl (100 mM) in
the incubation medium. These results indicate that, even if caspase-3 activation
is linked to the K(+) loss in cryopreserved PCLS, its inhibition does not allow
restoring the metabolic capacities. LiCl, acting on a target upstream of
caspase-3 inhibition, improves cell viability and allows glycogen accumulation
when added in culture medium of cryopreserved PCLS; and could thus be considered
as an interesting adjuvant in the culture of cryopreserved PCLS.

PMID: 16236333 [PubMed - in process]