Cryopreservation of Hepatocyte Microbeads for Clinical Transplantation
| Autor: | Anil Dhawan, Ragai R. Mitry, Celine Filippi, Leanne Glover, Robin D. Hughes, Suttiruk Jitraruch, Sharon C. Lehec |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Male Antioxidant medicine.medical_treatment Biomedical Engineering lcsh:Medicine cryopreservation Cryopreservation Andrology Rats Sprague-Dawley 03 medical and health sciences chemistry.chemical_compound cytoprotectants 0302 clinical medicine medicine clinical grade Animals Humans Viability assay Transplantation alginate encapsulation Dimethyl sulfoxide lcsh:R hepatocyte microbeads apoptosis Cell Biology Original Articles acute liver failure Liver Failure Acute Human serum albumin Rats Disease Models Animal 030104 developmental biology medicine.anatomical_structure chemistry Apoptosis Hepatocyte Hepatocytes 030211 gastroenterology & hepatology medicine.drug |
| Zdroj: | Cell Transplantation Cell Transplantation, Vol 26 (2017) Jitraruch, S, Dhawan, A, Hughes, R D, Filippi, C, Lehec, S C, Glover, L & Mitry, R R 2017, ' Cryopreservation of Hepatocyte Microbeads for Clinical Transplantation ', Cell Transplantation, vol. 26, no. 8, pp. 1341-1354 . https://doi.org/10.1177/0963689717720050 |
| ISSN: | 1555-3892 0963-6897 |
| Popis: | Intraperitoneal transplantation of hepatocyte microbeads is an attractive option for the management of acute liver failure. Encapsulation of hepatocytes in alginate microbeads supports their function and prevents immune attack of the cells. Establishment of banked cryopreserved hepatocyte microbeads is important for emergency use. The aim of this study was to develop an optimized protocol for cryopreservation of hepatocyte microbeads for clinical transplantation using modified freezing solutions. Four freezing solutions with potential for clinical application were investigated. Human and rat hepatocytes cryopreserved with University of Wisconsin (UW)/10% dimethyl sulfoxide (DMSO)/5% (300 mM) glucose and CryoStor CS10 showed better postthawing cell viability, attachment, and hepatocyte functions than with histidine–tryptophan–ketoglutarate/10% DMSO/5% glucose and Bambanker. The 2 freezing solutions that gave better results were studied with human and rat hepatocytes microbeads. Similar effects on cryopreserved microbead morphology (external and ultrastructural), viability, and hepatocyte-functions post thawing were observed over 7 d in culture. UW/DMSO/glucose, as a basal freezing medium, was used to investigate the additional effects of cytoprotectants: a pan-caspase inhibitor (benzyloxycarbonyl-Val-Ala-dl-Asp-fluoromethylketone [ZVAD]), an antioxidant (desferoxamine [DFO]), and a buffering and mechanical protectant (human serum albumin [HSA]) on RMBs. ZVAD (60 µM) had a beneficial effect on cell viability that was greater than with DFO (1 mM), HSA (2%), and basal freezing medium alone. Improvements in the ultrastructure of encapsulated hepatocytes and a lower degree of cell apoptosis were observed with all 3 cytoprotectants, with ZVAD tending to provide the greatest effect. Cytochrome P450 activity was significantly higher in the 3 cytoprotectant groups than with fresh microbeads. In conclusion, developing an optimized cryopreservation protocol by adding cytoprotectants such as ZVAD could improve the outcome of cryopreserved hepatocyte microbeads for future clinical use. |
| Databáze: | OpenAIRE |
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