Caspase inhibition reduces apoptotic death of cryopreserved porcine hepatocytes

Toshikazu Yagi, Joseph A. Hardin, Yunuen M. Valenzuela, Hideyuki Miyoshi, Gregory James Gores, Scott Nyberg

Research output: Contribution to journalArticle

99 Citations (Scopus)

Abstract

Cryopreserved porcine hepatocytes are a ready source of metabolic function for use in a bioartificial liver (BAL). However, cryopreservation is associated with a loss of hepatocyte viability. The mechanism of cell death during cryopreservation is incompletely understood, but may involve apoptosis through caspase activation. This study evaluates the cytoprotective effect of a global caspase inhibitor, benzyloxycarbonyl-Val-Ala-DL-Asp-fluoromethylketone (ZVADfmk) during cryopreservation of porcine hepatocytes. Freshly isolated porcine hepatocytes (viability, 97.4% ± 0.9%) were cryopreserved in 60 μmol/L ZVAD-fmk (+ZVAD group) or without ZVAD-fmk (-ZVAD group) for 24 to 72 hours. Apoptotic and necrotic death were both observed after thawing and after 24 hours of culture. Caspase 3-like activity was significantly reduced by ZVADfmk, and was associated with improved viability and reduced apoptotic death of porcine hepatocytes after cryopreservation. Mitochondrial membrane potential (MMP) was increased in cultures of porcine hepatocytes that were cryopreserved in ZVAD-fmk. These results demonstrate the following: 1) Caspase 3-like protease activation and apoptosis occurs in porcine hepatocytes during cryopreservation; and 2) mitochondrial injury in this process is reduced by caspase inhibition.

Original languageEnglish (US)
Pages (from-to)1432-1440
Number of pages9
JournalHepatology
Volume33
Issue number6
DOIs
StatePublished - 2001

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Caspases
Hepatocytes
Cryopreservation
Swine
Caspase 3
Artificial Liver
Apoptosis
Caspase Inhibitors
Mitochondrial Membrane Potential
Peptide Hydrolases
Cell Death
Wounds and Injuries

ASJC Scopus subject areas

  • Hepatology

Cite this

Caspase inhibition reduces apoptotic death of cryopreserved porcine hepatocytes. / Yagi, Toshikazu; Hardin, Joseph A.; Valenzuela, Yunuen M.; Miyoshi, Hideyuki; Gores, Gregory James; Nyberg, Scott.

In: Hepatology, Vol. 33, No. 6, 2001, p. 1432-1440.

Research output: Contribution to journalArticle

Yagi, Toshikazu ; Hardin, Joseph A. ; Valenzuela, Yunuen M. ; Miyoshi, Hideyuki ; Gores, Gregory James ; Nyberg, Scott. / Caspase inhibition reduces apoptotic death of cryopreserved porcine hepatocytes. In: Hepatology. 2001 ; Vol. 33, No. 6. pp. 1432-1440.
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