Engraftment and Albumin Production of Intrasplenically Transplanted Rat Hepatocytes (Sprague-Dawley), Freshly Isolated versus Cryopreserved, Into Nagase Analbuminemic Rats (NAR)
Banking of cryopreserved hepatocytes is a prerequisite for large-scale hepatocyte transplantation in the clinic. We compared the efficacy of intrasplenic transplantation into Nagase analbuminemic rats (NAR) of freshly isolated (FIH) and cryopreserved (CH) hepatocytes. Hepatocytes were cryopreserved...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2001-01-01
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| Series: | Cell Transplantation |
| Online Access: | https://doi.org/10.3727/000000001783987034 |
| _version_ | 1828860869615812608 |
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| author | Pascale David Eliane Alexandre Maxime Audet Marie-Pierre Chenard-Neu Philippe Wolf Daniel Jaeck Agnès Azimzadeh Lysiane Richert |
| author_facet | Pascale David Eliane Alexandre Maxime Audet Marie-Pierre Chenard-Neu Philippe Wolf Daniel Jaeck Agnès Azimzadeh Lysiane Richert |
| author_sort | Pascale David |
| collection | DOAJ |
| description | Banking of cryopreserved hepatocytes is a prerequisite for large-scale hepatocyte transplantation in the clinic. We compared the efficacy of intrasplenic transplantation into Nagase analbuminemic rats (NAR) of freshly isolated (FIH) and cryopreserved (CH) hepatocytes. Hepatocytes were cryopreserved using a controlled rate freezing protocol. Albumin production of thawed CH and FIH was measured in vitro in culture by ELISA and by Western blot. After in vivo intrasplenic transplantation of NAR with either FIH or CH we assessed 1) albumin in the serum of recipients by ELISA and by Western blotting analysis at different time intervals, and 2) hepatocyte engraftment by albumin immunohistochemical staining into spleens and livers at euthanasia. In vitro, albumin was produced up to day 4 of culture in both CH and FIH. In vivo, no intrasplenic engraftment of hepatocytes occurred. Intrahepatic engraftment of CH (cell number/mm2) was significantly (twofold) lower than that of FIH and appeared only as isolated cells and small (<10 cells) clusters, while bigger clusters (>10 cells) were observed with FIH. In the FIH group, serum albumin production was observed up to 32 – 49 days posttransplantation while in the CH group no serum albumin production was detected. Our results emphasize the need to improve 1) hepatocyte transplantation procedures either by repeated hepatocytes injections and/or by transplantation under a regeneration response, and 2) the freeze/thaw protocols of hepatocytes. |
| first_indexed | 2024-12-13T02:49:00Z |
| format | Article |
| id | doaj.art-66be2f1a009a40b0af4aedb16895d2ff |
| institution | Directory Open Access Journal |
| issn | 0963-6897 1555-3892 |
| language | English |
| last_indexed | 2024-12-13T02:49:00Z |
| publishDate | 2001-01-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Cell Transplantation |
| spelling | doaj.art-66be2f1a009a40b0af4aedb16895d2ff2022-12-22T00:02:06ZengSAGE PublishingCell Transplantation0963-68971555-38922001-01-011010.3727/000000001783987034Engraftment and Albumin Production of Intrasplenically Transplanted Rat Hepatocytes (Sprague-Dawley), Freshly Isolated versus Cryopreserved, Into Nagase Analbuminemic Rats (NAR)Pascale David0Eliane Alexandre1Maxime Audet2Marie-Pierre Chenard-Neu3Philippe Wolf4Daniel Jaeck5Agnès Azimzadeh6Lysiane Richert7Laboratoire de Chirurgie Expérimentale, Fondation Transplantation, 5, avenue Molière, 67200 Strasbourg, FranceLaboratoire de Chirurgie Expérimentale, Fondation Transplantation, 5, avenue Molière, 67200 Strasbourg, FranceCentre de Chirurgie Viscérale et de Transplantation, Hôpital de Hautepierre, Avenue Molière, 67098 Strasbourg, FranceService d'Anatomo-Pathologie, Hôpital de Hautepierre, Avenue Molière, 67098 Strasbourg, FranceCentre de Chirurgie Viscérale et de Transplantation, Hôpital de Hautepierre, Avenue Molière, 67098 Strasbourg, FranceCentre de Chirurgie Viscérale et de Transplantation, Hôpital de Hautepierre, Avenue Molière, 67098 Strasbourg, FranceLaboratoire de Chirurgie Expérimentale, Fondation Transplantation, 5, avenue Molière, 67200 Strasbourg, FranceLaboratoire de Biologie Cellulaire, Faculté de Pharmacie, 4, Place Saint Jacques, 25030 Besançon, FranceBanking of cryopreserved hepatocytes is a prerequisite for large-scale hepatocyte transplantation in the clinic. We compared the efficacy of intrasplenic transplantation into Nagase analbuminemic rats (NAR) of freshly isolated (FIH) and cryopreserved (CH) hepatocytes. Hepatocytes were cryopreserved using a controlled rate freezing protocol. Albumin production of thawed CH and FIH was measured in vitro in culture by ELISA and by Western blot. After in vivo intrasplenic transplantation of NAR with either FIH or CH we assessed 1) albumin in the serum of recipients by ELISA and by Western blotting analysis at different time intervals, and 2) hepatocyte engraftment by albumin immunohistochemical staining into spleens and livers at euthanasia. In vitro, albumin was produced up to day 4 of culture in both CH and FIH. In vivo, no intrasplenic engraftment of hepatocytes occurred. Intrahepatic engraftment of CH (cell number/mm2) was significantly (twofold) lower than that of FIH and appeared only as isolated cells and small (<10 cells) clusters, while bigger clusters (>10 cells) were observed with FIH. In the FIH group, serum albumin production was observed up to 32 – 49 days posttransplantation while in the CH group no serum albumin production was detected. Our results emphasize the need to improve 1) hepatocyte transplantation procedures either by repeated hepatocytes injections and/or by transplantation under a regeneration response, and 2) the freeze/thaw protocols of hepatocytes.https://doi.org/10.3727/000000001783987034 |
| spellingShingle | Pascale David Eliane Alexandre Maxime Audet Marie-Pierre Chenard-Neu Philippe Wolf Daniel Jaeck Agnès Azimzadeh Lysiane Richert Engraftment and Albumin Production of Intrasplenically Transplanted Rat Hepatocytes (Sprague-Dawley), Freshly Isolated versus Cryopreserved, Into Nagase Analbuminemic Rats (NAR) Cell Transplantation |
| title | Engraftment and Albumin Production of Intrasplenically Transplanted Rat Hepatocytes (Sprague-Dawley), Freshly Isolated versus Cryopreserved, Into Nagase Analbuminemic Rats (NAR) |
| title_full | Engraftment and Albumin Production of Intrasplenically Transplanted Rat Hepatocytes (Sprague-Dawley), Freshly Isolated versus Cryopreserved, Into Nagase Analbuminemic Rats (NAR) |
| title_fullStr | Engraftment and Albumin Production of Intrasplenically Transplanted Rat Hepatocytes (Sprague-Dawley), Freshly Isolated versus Cryopreserved, Into Nagase Analbuminemic Rats (NAR) |
| title_full_unstemmed | Engraftment and Albumin Production of Intrasplenically Transplanted Rat Hepatocytes (Sprague-Dawley), Freshly Isolated versus Cryopreserved, Into Nagase Analbuminemic Rats (NAR) |
| title_short | Engraftment and Albumin Production of Intrasplenically Transplanted Rat Hepatocytes (Sprague-Dawley), Freshly Isolated versus Cryopreserved, Into Nagase Analbuminemic Rats (NAR) |
| title_sort | engraftment and albumin production of intrasplenically transplanted rat hepatocytes sprague dawley freshly isolated versus cryopreserved into nagase analbuminemic rats nar |
| url | https://doi.org/10.3727/000000001783987034 |
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