| Cryopreservation of Encapsulated Liver Spheroids for a Bioartificial Liver: Reducing Latent Cryoinjury using an Ice Nucleating Agent. | |
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MedLine Citation:
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PMID: 21410301 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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Introduction: Acute liver failure has high mortality due to donor organ shortages. A bioartificial liver could "bridge the gap" to transplant or spontaneous recovery. Alginate encapsulation of HepG2 cells enables cell spheroid formation providing sufficient functional biomass. Cryopreservation of these spheroids would allow an off-the-shelf capability for unpredictable emergency use. Cell death during cryopreservation often results from intracellular ice formation, following supercooling. An ice nucleating agent (INA), crystalline cholesterol, was trialled to reduce supercooling and subsequent cryoinjury. Materials & Methods: Spheroids were cooled in a controlled rate freezer in 12% dimethylsulfoxide/Celsior +/- INA and sample temperatures recorded throughout. Viability was assessed using fluorescent staining with image analysis, cell number by nuclei count, function using assays to detect liver-specific protein synthesis and secretion, MTT reduction and broad-spectrum cytochrome P450 activity and apoptosis estimated using a luminescence-based techniques. Results: Spheroids cryopreserved without INA displayed latent cryoinjury in the first 6 hours following thawing. INA reduced supercooling during cryopreservation and also latent cryoinjury. Cell numbers, viability and function as measured over 72 hours post-thaw were all improved when INA was present during cryopreservation. |
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Authors:
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Isobel Massie; Clare Selden; Humphrey Hodgson; Barry Fuller |
Publication Detail:
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Type: JOURNAL ARTICLE Date: 2011-3-16 |
Journal Detail:
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Title: Tissue engineering. Part C, Methods Volume: - ISSN: 1937-3392 ISO Abbreviation: - Publication Date: 2011 Mar |
Date Detail:
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Created Date: 2011-3-17 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101466663 Medline TA: Tissue Eng Part C Methods Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Affiliation:
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UCL, Centre for Hepatology, London, United Kingdom; i.massie@medsch.ucl.ac.uk. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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