Document Detail


The application of 3D micropatterning of agarose substrate for cell culture and in situ comet assays.
MedLine Citation:
PMID:  20149429     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We report the fabrication of a 3D micropatterned agarose substrate that enables the culture of single or multiple cells. Patterning was performed on dried agarose using deep UV irradiation leading to 6-microm-deep micropatterns of 25-70 microm in diameter. Cell adhesion was facilitated by the specific grafting of ECM (extra cellular matrix) proteins such as fibronectin into the micropatterns. We show that the pattern size induced the adhesion of one or more cells, thus allowing precise control of the cell number used in the assay, and that cells proliferated similarly as in standard culture conditions. Moreover, cell polarity appeared well preserved on this substrate, so polarized cells like hepatoma HepaRG cells might maintain their differentiation status and act as primary human hepatocytes for hepatotoxicity testing. These 3D patterned culture slides have been successfully used for in situ comet assays and there is evidence that the genotoxic effects of sub-cytotoxic concentrations of drugs could be analyzed in a large number of single HeLa cells. Coupled with the parallel-based design of the 3D micropatterning, which allows automated image analysis, these results strongly indicate that this new cell array system is suitable for high-throughput cytotoxicity and genotoxicity screening applications.
Authors:
Emilie Mercey; Patricia Obe?d; Denise Glaise; Maria-Luisa Calvo-Mu?oz; Christiane Guguen-Guillouzo; Brigitte Fouqu?
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-02-10
Journal Detail:
Title:  Biomaterials     Volume:  31     ISSN:  1878-5905     ISO Abbreviation:  Biomaterials     Publication Date:  2010 Apr 
Date Detail:
Created Date:  2010-02-23     Completed Date:  2010-06-03     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8100316     Medline TA:  Biomaterials     Country:  England    
Other Details:
Languages:  eng     Pagination:  3156-65     Citation Subset:  IM    
Copyright Information:
Copyright 2010 Elsevier Ltd. All rights reserved.
Affiliation:
CEA, DSV, IRTSV, Laboratoire Biopuces, Grenoble, France.
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MeSH Terms
Descriptor/Qualifier:
Cell Line, Tumor
Comet Assay*
Extracellular Matrix Proteins / chemistry
Fluorescent Dyes
Humans
Microscopy, Confocal
Sepharose / chemistry*
Ultraviolet Rays
Chemical
Reg. No./Substance:
0/Extracellular Matrix Proteins; 0/Fluorescent Dyes; 9012-36-6/Sepharose

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


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