Cell-culturing substrates where cell adhesion can be switched on by external stimuli during cell cultivation are useful scaffolds for tissue engineering, cell-based drug screening, and fundamental cellular studies. Here, we show a new strategy for photoactivation of a substrate for cell adhesion under standard fluorescence microscopes. A glass substrate chemically modified with an alkylsiloxane having a photocleavable 2-nitrobenzyl group was coated with bovine serum albumin to prevent cell adhesion. Upon irradiation under a fluorescence microscope, the protein was replaced with fibronectin, which made the irradiated region cell-adhesive. Subsequent seeding of HEK293 or COS7 cells produced patterns corresponding to the irradiated patterns. We succeeded for the first time in positioning single cells in proximity to cultivating single cells. The present method provides a general strategy for positioning single cells of same or different types at any locations on the substrate and will be useful for studying cell-cell interactions.
Spatiotemporal control of cell migration was achieved on a photoactivatable cell-culturing substrate. Single cells were micropatterned on the substrate and were induced to extend protrusions led by lamellipodia or filopodia alternatively by the subsequent formation of wide or narrow paths in their surroundings, respectively. By tracking the migration of single cells in a microarray format, we performed quantitative analysis of the migration rates of single cells.
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