Versatile gradients of covalently bound proteins on microstructured substrates.
In this work, we propose an easy method to produce highly tunable gradients of covalently bound proteins on topographically modified poly(methyl methacrylate). We used a microfluidic approach to obtain linear gradients with high slope (0.5 pmol·cm(-2)·mm(-1)), relevant at the single-cell level. These protein gradients were characterized using fluorescence microscopy and surface plasmon resonance. Both experimental results and theoretical modeling on the protein gradients generated have proved them to be highly reproducible, stable up to 7 days, and easily tunable. This method enables  formation of versatile cell culture platforms combining both complex biochemical and physical cues in an attempt to approach in vitro cell culture methods to in vivo cellular microenvironments.
Institute for Bioengineering of Catalonia, C/Baldiri Reixac 11-15, 08028 Barcelona, Spain. jcomelles@ibecbarcelona.eu
Jordi
J
Comelles
Verónica
V
Hortigüela
Josep
J
Samitier
Elena
E
Martínez
D000818:Animals; D002448:Cell Adhesion; D051379:Mice; D046210:Microfluidic Analytical Techniques; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D019904:Polymethyl Methacrylate / Q000737:chemistry*; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D020349:Surface Plasmon Resonance
D011506:Proteins; D019904:Polymethyl Methacrylate