Apoptotic Cell Death and Function of Cryopreserved Porcine Hepatocytes in a Bioartificial Liver
We have previously shown that cryopreservation leads to increased apoptotic death of porcine hepatocytes intended for use in a bioartificial liver (BAL). This study was designed to determine if a broad-spectrum caspase inhibitor, IDN–1965, reduced apoptosis and increased function of cryopreserved po...
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doaj-252717a4f37b445ca7bb71e381becc752020-11-25T03:22:47ZengSAGE PublishingCell Transplantation0963-68971555-38922003-03-011210.3727/000000003108746696Apoptotic Cell Death and Function of Cryopreserved Porcine Hepatocytes in a Bioartificial LiverTakakazu Matsushita0Toshikazu Yagi1Joseph A. Hardin2Jennifer D. Cragun3Frank W. Crow4H. Robert Bergen5Gregory J. Gores6Scott L. Nyberg M.D., Ph.D.7Division of Transplantation SurgeryDivision of Transplantation SurgeryDivision of Transplantation SurgeryDivision of Biochemistry and Molecular BiologyDivision of Biochemistry and Molecular BiologyDivision of Biochemistry and Molecular BiologyDivision of Gastroenterology and Hepatology, Mayo Clinic, Rochester, MNDivision of Transplantation SurgeryWe have previously shown that cryopreservation leads to increased apoptotic death of porcine hepatocytes intended for use in a bioartificial liver (BAL). This study was designed to determine if a broad-spectrum caspase inhibitor, IDN–1965, reduced apoptosis and increased function of cryopreserved porcine hepatocytes in static culture or in a BAL. Porcine hepatocytes were studied immediately after isolation and after 2 weeks of cryopreservation in liquid nitrogen using medium supplemented with 25 μmol/L IDN-1965 or vehicle. Both apoptotic and necrotic cells were observed in cultures of fresh and cryopreserved hepatocytes, but the percentage of apoptotic cells increased after cryopreservation. Cryopreservation in IDN-1965 improved hepatocyte viability and reduced apoptotic cell death determined by TUNEL assay. Cryopreservation of hepatocytes in IDN-1965 was also associated with reduced caspase 3-like activity, decreased release of cytochrome c from mitochondria, and a slower decline in mitochondrial membrane potential after thawing. These markers of apoptosis were lowest after cryopreservation when IDN-1965 was added to both the culture and cryopreservation medium. Functional markers of hepatocyte activity (albumin production, diazepam metabolism, urea production) were also increased after cryopreservation and culture of hepatocytes in medium supplemented with 25 μmol/L IDN-1965. Cryopreservation of porcine hepatocytes in the presence of caspase inhibitor IDN-1965 was associated with reduced apoptosis and improved function of porcine hepatocytes in both static culture and a perfused BAL. These data demonstrate that inhibition of apoptosis also preserves cell function.https://doi.org/10.3727/000000003108746696 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Takakazu Matsushita Toshikazu Yagi Joseph A. Hardin Jennifer D. Cragun Frank W. Crow H. Robert Bergen Gregory J. Gores Scott L. Nyberg M.D., Ph.D. |
spellingShingle |
Takakazu Matsushita Toshikazu Yagi Joseph A. Hardin Jennifer D. Cragun Frank W. Crow H. Robert Bergen Gregory J. Gores Scott L. Nyberg M.D., Ph.D. Apoptotic Cell Death and Function of Cryopreserved Porcine Hepatocytes in a Bioartificial Liver Cell Transplantation |
author_facet |
Takakazu Matsushita Toshikazu Yagi Joseph A. Hardin Jennifer D. Cragun Frank W. Crow H. Robert Bergen Gregory J. Gores Scott L. Nyberg M.D., Ph.D. |
author_sort |
Takakazu Matsushita |
title |
Apoptotic Cell Death and Function of Cryopreserved Porcine Hepatocytes in a Bioartificial Liver |
title_short |
Apoptotic Cell Death and Function of Cryopreserved Porcine Hepatocytes in a Bioartificial Liver |
title_full |
Apoptotic Cell Death and Function of Cryopreserved Porcine Hepatocytes in a Bioartificial Liver |
title_fullStr |
Apoptotic Cell Death and Function of Cryopreserved Porcine Hepatocytes in a Bioartificial Liver |
title_full_unstemmed |
Apoptotic Cell Death and Function of Cryopreserved Porcine Hepatocytes in a Bioartificial Liver |
title_sort |
apoptotic cell death and function of cryopreserved porcine hepatocytes in a bioartificial liver |
publisher |
SAGE Publishing |
series |
Cell Transplantation |
issn |
0963-6897 1555-3892 |
publishDate |
2003-03-01 |
description |
We have previously shown that cryopreservation leads to increased apoptotic death of porcine hepatocytes intended for use in a bioartificial liver (BAL). This study was designed to determine if a broad-spectrum caspase inhibitor, IDN–1965, reduced apoptosis and increased function of cryopreserved porcine hepatocytes in static culture or in a BAL. Porcine hepatocytes were studied immediately after isolation and after 2 weeks of cryopreservation in liquid nitrogen using medium supplemented with 25 μmol/L IDN-1965 or vehicle. Both apoptotic and necrotic cells were observed in cultures of fresh and cryopreserved hepatocytes, but the percentage of apoptotic cells increased after cryopreservation. Cryopreservation in IDN-1965 improved hepatocyte viability and reduced apoptotic cell death determined by TUNEL assay. Cryopreservation of hepatocytes in IDN-1965 was also associated with reduced caspase 3-like activity, decreased release of cytochrome c from mitochondria, and a slower decline in mitochondrial membrane potential after thawing. These markers of apoptosis were lowest after cryopreservation when IDN-1965 was added to both the culture and cryopreservation medium. Functional markers of hepatocyte activity (albumin production, diazepam metabolism, urea production) were also increased after cryopreservation and culture of hepatocytes in medium supplemented with 25 μmol/L IDN-1965. Cryopreservation of porcine hepatocytes in the presence of caspase inhibitor IDN-1965 was associated with reduced apoptosis and improved function of porcine hepatocytes in both static culture and a perfused BAL. These data demonstrate that inhibition of apoptosis also preserves cell function. |
url |
https://doi.org/10.3727/000000003108746696 |
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