Fabrication of a cell-adhesive microwell array for 3-dimensional in vitro cell model
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Microarrays of single living cells facilitate elucidation of the biological effects of a drug on a single cell providing large amounts of information regarding cellular chemistry at the molecular level in quantitative manner. Compared to 2D cultures, 3D in vitro cell models provide a more realistic cellular environment and permit the reproduction of in vitro cellular phenotypes.
We fabricated a microwell array for single cell analysis that could preserve the native morphology and functionality of HeLa cells by creating microwells with a cell-adhesive inner surface. Dopaminergic inorganic-organic hybrid resin (HR4-DOPA) was used for cell-adhesive layer and fabricated by spin-coating and micro contact printing methods. The physical dimensions of the microwells were designed to achieve high single-cell occupancy.
Microwells was 25 × 25 μm2 in size from the diameter of a HeLa cell. An optimal depth of microwells and cell concentration for HeLa single-cell occupancy was investigated; 25-μm-deep microwells at 1.0 × 106 cells/ml demonstrated the highest single-cell occupancy of 67.5% while providing a sufficient cell-adhesive surface area for long-term cell culture. We obtained singly occupied microwells with HeLa cells in 1072 microwells from 8 microwell arrays in the 2 × 2mm2 area and with only 5.9% array-to-array variation.
The number of singly occupied microwells and array-to-array variation provides adequate throughput for accurate quantification in advanced single-cell analysis.
KeywordsMicrowell array 3-Dimensional in vitro cell model Cell adhesive surface Single cell occupancy HeLa cell
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