Document Detail

Storage temperatures for cold-chain delivery in cell therapy: a study of alginate-encapsulated liver cell spheroids stored at -80°c or -170°c for up to 1 year.
MedLine Citation:
PMID:  22834979     Owner:  NLM     Status:  MEDLINE    
INTRODUCTION: A bioartificial liver comprising alginate-encapsulated liver cell spheroids (ELS) could bridge the gap to transplant or spontaneous recovery in acute liver failure, but will be required for emergency use, necessitating cryopreservation. A cryopreservation protocol has been developed, but beyond this, the feasibility of cold-chain storage is considered here. Cryopreservation will be increasingly required for timely delivery of tissue and bioengineered products, and significant, but often, over-looked factors that impact on cost and ease of clinical application are the storage temperature and useful preservation time. Storage in the vapor phase of liquid nitrogen (∼-170°C) is the gold standard, but for safety and economic purposes, storing ELS in electric freezers at -80°C may be preferable.
METHODS: ELS were cryopreserved using an optimized protocol and stored at either -80°C or at -170°C for up to 1 year. ELS were removed from storage after 1, 2, 3, 6, 9, or 12 months, and recovery was assessed 24 h postwarming. Cell recovery was assessed using viability (fluorescent staining with image analysis), cell number (nuclei count), and functional (hepatospecific protein enzyme-linked immunosorbent assay) assays.
RESULTS: Viability, the viable cell number, and function of ELS stored at -170°C were maintained at similar values throughout the year. In contrast, ELS stored at -80°C exhibited decreased viability, viable cell numbers, and function by as early as 1 month. Progressive deterioration was subsequently observed. After 12 months of storage at -80°C, viable cell recovery of ELS was ∼15% that of ELS stored at -170°C.
CONCLUSIONS: While convenience and cost might support the use of -80°C for storage of multicellular bioengineered products such as ELS, results indicate rapid deterioration in functional recoveries after only a few weeks. This study demonstrates that storage temperature is an important consideration in regenerative medicine and caution should be applied by limiting storage at -80°C to only a few weeks.
Isobel Massie; Clare Selden; Humphrey Hodgson; Barry Fuller
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-09-07
Journal Detail:
Title:  Tissue engineering. Part C, Methods     Volume:  19     ISSN:  1937-3392     ISO Abbreviation:  Tissue Eng Part C Methods     Publication Date:  2013 Mar 
Date Detail:
Created Date:  2013-01-29     Completed Date:  2013-07-11     Revised Date:  2014-03-09    
Medline Journal Info:
Nlm Unique ID:  101466663     Medline TA:  Tissue Eng Part C Methods     Country:  United States    
Other Details:
Languages:  eng     Pagination:  189-95     Citation Subset:  IM    
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MeSH Terms
Cell Survival
Coated Materials, Biocompatible / chemistry*
Cold Temperature
Cryopreservation / methods*
Hep G2 Cells
Hepatocytes / cytology*,  physiology
Liver, Artificial*
Organ Preservation / methods*
Spheroids, Cellular / cytology*,  physiology
Grant Support
066327//Wellcome Trust; MR/K500720/1//Medical Research Council
Reg. No./Substance:
0/Coated Materials, Biocompatible

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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