Document Detail


Cryoprotectant delivery and removal from murine insulinomas at vitrification-relevant concentrations.
MedLine Citation:
PMID:  17533114     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Development of optimal cryopreservation protocols requires delivery and removal of cryoprotective agents (CPAs) in such a way that negative osmotic and cytotoxic effects on cells are minimized. This is especially true for vitrification, where high CPA concentrations are employed. In this study, we report on the determination of cell membrane permeability parameters for water (L(p)) and solute (P(s)), and on the design and experimental verification of CPA addition and removal protocols at vitrification-relevant concentrations for a murine insulinoma cell line, betaTC-tet cells. Using membrane permeability values and osmotic tolerance limits, mathematical modeling and computer simulations were used to design CPA addition and removal protocols at high concentrations. The cytotoxic effects of CPAs were also evaluated. Cells were able to tolerate the addition and removal of 2.5M dimethyl sulfoxide (DMSO) and 2.5M 1,2 propanediol (PD) in single steps, but required multi-step addition and removal with 3.0M DMSO, 3.0M PD, and a vitrification-relevant concentration of 3.0M DMSO+3.0M PD. Cytotoxicity studies revealed that betaTC-tet cells were able to tolerate the presence of single component 6.0M DMSO and 6.0M PD and to a lesser extent 3.0M DMSO+3.0M PD. These results determine the time and concentration domain of CPA exposure that cells can tolerate and are essential for designing cryopreservation protocols for free cells as well as cells in engineered tissues.
Authors:
Indra Neil Mukherjee; Ying C Song; Athanassios Sambanis
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2007-04-10
Journal Detail:
Title:  Cryobiology     Volume:  55     ISSN:  0011-2240     ISO Abbreviation:  Cryobiology     Publication Date:  2007 Aug 
Date Detail:
Created Date:  2007-07-30     Completed Date:  2007-12-27     Revised Date:  2014-09-14    
Medline Journal Info:
Nlm Unique ID:  0006252     Medline TA:  Cryobiology     Country:  United States    
Other Details:
Languages:  eng     Pagination:  10-8     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Line
Cell Membrane Permeability*
Cryopreservation / methods*
Cryoprotective Agents / metabolism*,  toxicity
Insulinoma / metabolism*
Mice
Osmotic Pressure
Grant Support
ID/Acronym/Agency:
R01 DK073991/DK/NIDDK NIH HHS; R01 DK073991-03/DK/NIDDK NIH HHS; R01DK073991/DK/NIDDK NIH HHS; R43 DK062533-01/DK/NIDDK NIH HHS; R43DK062533/DK/NIDDK NIH HHS
Chemical
Reg. No./Substance:
0/Cryoprotective Agents
Comments/Corrections

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