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The application of 3D micropatterning of agarose substrate for cell culture and in situ comet assays

Paper ID Volume ID Publish Year Pages File Format Full-Text
8120 578 2010 10 PDF Available
Title
The application of 3D micropatterning of agarose substrate for cell culture and in situ comet assays
Abstract

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-μm-deep micropatterns of 25–70 μm 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.

Keywords
3D micropatterning; Agarose; Cell array; Comet assay; HepaRG hepatocytes; Genotoxicity
First Page Preview
The application of 3D micropatterning of agarose substrate for cell culture and in situ comet assays
Publisher
Database: Elsevier - ScienceDirect
Journal: Biomaterials - Volume 31, Issue 12, April 2010, Pages 3156–3165
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering