Cryopreservation of Human Hepatocytes Alters the Mitochondrial Respiratory Chain Complex 1
Transplantation of human hepatocytes has recently been demonstrated as a safe alternative to partially correct liver inborn errors of metabolism. Cryopreservation remains the most appropriate way of cell banking. However, mitochondrial-mediated apoptosis has been reported after cryopreservation and...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2007-04-01
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| Series: | Cell Transplantation |
| Online Access: | https://doi.org/10.3727/000000007783464821 |
| _version_ | 1818922946289205248 |
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| author | Xavier Stéphenne Mustapha Najimi Dung Khuu Ngoc Françoise Smets Louis Hue Bruno Guigas Etienne M. Sokal |
| author_facet | Xavier Stéphenne Mustapha Najimi Dung Khuu Ngoc Françoise Smets Louis Hue Bruno Guigas Etienne M. Sokal |
| author_sort | Xavier Stéphenne |
| collection | DOAJ |
| description | Transplantation of human hepatocytes has recently been demonstrated as a safe alternative to partially correct liver inborn errors of metabolism. Cryopreservation remains the most appropriate way of cell banking. However, mitochondrial-mediated apoptosis has been reported after cryopreservation and little is known on the involved molecular mechanisms. The aim of this study was to investigate mitochondrial functions of freshly isolated and cryopreserved/thawed hepatocytes from mice and humans. We report here that cryopreservation induced a dramatic drop of ATP levels in hepatocytes. The oxygen consumption rate of cryopreserved/thawed hepatocytes was significantly lower compared to fresh cells. In addition, the uncoupling effect of 2,4-dinitrophenol was lost, in parallel with a reduction of mitochondrial membrane potential. Furthermore, a decrease in mitochondrial respiratory rate was evidenced on permeabilized hepatocytes in the presence of substrate for the respiratory chain complex 1. Interestingly, this effect was less marked with a substrate for complex 2. Electron microscopy examination indicated that mitochondria were swollen and devoid of cristae after cryopreservation. These changes could explain the cytosolic release of the proapoptotic protein cytochrome c in cryopreserved cells. Nevertheless, no caspase 9-3 activation and only few apoptotic and necrotic cells were found, indicating that the subsequent cell death program was not yet evidenced. Our results demonstrate that cryopreservation of hepatocytes induced alteration of the mitochondrial machinery. They also suggest that, in addition to technical progress in the cryopreservation procedure, protection of the respiratory chain complex 1 should be considered to improve the quality of cryopreserved hepatocytes. |
| first_indexed | 2024-12-20T02:01:37Z |
| format | Article |
| id | doaj.art-998bc8f3dba14e33ae93b7dea7e3babe |
| institution | Directory Open Access Journal |
| issn | 0963-6897 1555-3892 |
| language | English |
| last_indexed | 2024-12-20T02:01:37Z |
| publishDate | 2007-04-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Cell Transplantation |
| spelling | doaj.art-998bc8f3dba14e33ae93b7dea7e3babe2022-12-21T19:57:18ZengSAGE PublishingCell Transplantation0963-68971555-38922007-04-011610.3727/000000007783464821Cryopreservation of Human Hepatocytes Alters the Mitochondrial Respiratory Chain Complex 1Xavier Stéphenne0Mustapha Najimi1Dung Khuu Ngoc2Françoise Smets3Louis Hue4Bruno Guigas5Etienne M. Sokal6 Laboratory of Paediatric Hepatology and Cell Therapy, Université catholique de Louvain and Cliniques St Luc, Brussels, Belgium Laboratory of Paediatric Hepatology and Cell Therapy, Université catholique de Louvain and Cliniques St Luc, Brussels, Belgium Laboratory of Paediatric Hepatology and Cell Therapy, Université catholique de Louvain and Cliniques St Luc, Brussels, Belgium Laboratory of Paediatric Hepatology and Cell Therapy, Université catholique de Louvain and Cliniques St Luc, Brussels, Belgium Hormone and Metabolic Research Unit, School of Medicine, Université catholique de Louvain and Institute of Cellular Pathology, Brussels, Belgium Hormone and Metabolic Research Unit, School of Medicine, Université catholique de Louvain and Institute of Cellular Pathology, Brussels, Belgium Laboratory of Paediatric Hepatology and Cell Therapy, Université catholique de Louvain and Cliniques St Luc, Brussels, BelgiumTransplantation of human hepatocytes has recently been demonstrated as a safe alternative to partially correct liver inborn errors of metabolism. Cryopreservation remains the most appropriate way of cell banking. However, mitochondrial-mediated apoptosis has been reported after cryopreservation and little is known on the involved molecular mechanisms. The aim of this study was to investigate mitochondrial functions of freshly isolated and cryopreserved/thawed hepatocytes from mice and humans. We report here that cryopreservation induced a dramatic drop of ATP levels in hepatocytes. The oxygen consumption rate of cryopreserved/thawed hepatocytes was significantly lower compared to fresh cells. In addition, the uncoupling effect of 2,4-dinitrophenol was lost, in parallel with a reduction of mitochondrial membrane potential. Furthermore, a decrease in mitochondrial respiratory rate was evidenced on permeabilized hepatocytes in the presence of substrate for the respiratory chain complex 1. Interestingly, this effect was less marked with a substrate for complex 2. Electron microscopy examination indicated that mitochondria were swollen and devoid of cristae after cryopreservation. These changes could explain the cytosolic release of the proapoptotic protein cytochrome c in cryopreserved cells. Nevertheless, no caspase 9-3 activation and only few apoptotic and necrotic cells were found, indicating that the subsequent cell death program was not yet evidenced. Our results demonstrate that cryopreservation of hepatocytes induced alteration of the mitochondrial machinery. They also suggest that, in addition to technical progress in the cryopreservation procedure, protection of the respiratory chain complex 1 should be considered to improve the quality of cryopreserved hepatocytes.https://doi.org/10.3727/000000007783464821 |
| spellingShingle | Xavier Stéphenne Mustapha Najimi Dung Khuu Ngoc Françoise Smets Louis Hue Bruno Guigas Etienne M. Sokal Cryopreservation of Human Hepatocytes Alters the Mitochondrial Respiratory Chain Complex 1 Cell Transplantation |
| title | Cryopreservation of Human Hepatocytes Alters the Mitochondrial Respiratory Chain Complex 1 |
| title_full | Cryopreservation of Human Hepatocytes Alters the Mitochondrial Respiratory Chain Complex 1 |
| title_fullStr | Cryopreservation of Human Hepatocytes Alters the Mitochondrial Respiratory Chain Complex 1 |
| title_full_unstemmed | Cryopreservation of Human Hepatocytes Alters the Mitochondrial Respiratory Chain Complex 1 |
| title_short | Cryopreservation of Human Hepatocytes Alters the Mitochondrial Respiratory Chain Complex 1 |
| title_sort | cryopreservation of human hepatocytes alters the mitochondrial respiratory chain complex 1 |
| url | https://doi.org/10.3727/000000007783464821 |
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