Document Detail


Cryopreservation of adherent cells: strategies to improve cell viability and function after thawing.
MedLine Citation:
PMID:  19196129     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The commonly applied cryopreservation protocols routinely used in laboratories worldwide were developed for simple cell suspensions, and their application to complex systems, such as cell monolayers, tissues, or biosynthetic constructs, is not straightforward. In particular for monolayer cultures, cell detachment and membrane damage are often observed after cryopreservation. In this work, combined strategies for the cryopreservation of cells attached to Matrigel-coated well plate's surfaces were investigated based on cell entrapment in clinicalgrade, ultra-high viscosity alginate using two cell lines, neuroblastoma N2a and colon adenocarcinoma Caco-2, with distinct structural and functional characteristics. As the cryopreservation medium, serum-free CryoStor solution was compared with serum-supplemented culture medium, both containing 10% DMSO. Using culture medium, entrapment beneath an alginate layer was needed to improve cell recovery by minimizing membrane damage and cell detachment after thawing; nevertheless, up to 50% cell death still occurred within 24 h after thawing. The use of CryoStor solution represented a considerable improvement of the cryopreservation process for both cell lines, allowing the maintenance of high postthaw membrane integrity as well as full recovery of metabolic activity and differentiation capacity within 24 h postthawing; in this case, entrapment beneath an alginate layer did not confer further protection to cryopreserved Caco-2 cells, but was crucial for maintenance of attachment and integrity of N2a neuronal networks.
Authors:
Rita Malpique; Friederike Ehrhart; Alisa Katsen-Globa; Heiko Zimmermann; Paula M Alves
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Tissue engineering. Part C, Methods     Volume:  15     ISSN:  1937-3392     ISO Abbreviation:  -     Publication Date:  2009 Sep 
Date Detail:
Created Date:  2009-09-03     Completed Date:  2009-11-13     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101466663     Medline TA:  Tissue Eng Part C Methods     Country:  United States    
Other Details:
Languages:  eng     Pagination:  373-86     Citation Subset:  IM    
Affiliation:
Animal Cell Technology, IBET/ITQB-UNL, 27801-901 Oeiras, Portugal.
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MeSH Terms
Descriptor/Qualifier:
Caco-2 Cells
Cell Adhesion / drug effects*
Cell Survival / drug effects
Cryopreservation / methods*
Cryoprotective Agents / pharmacology*
Cytoprotection / physiology
Heating
Humans
Chemical
Reg. No./Substance:
0/Cryoprotective Agents

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


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