Self-Patterning of PC12 Cells on Micro-Structures of Protein-Modified Particles
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Micro-patterning of nerve cells is important technical issue to clarify their function and/or apply cell-based biosensors. Self-assembled particles are candidate for simply micro-structured scaffold to enable cells to adhere selectively. In this present study, using silica (SiO2) particles modified by specific protein, effects of particle-structure and protein-modification are investigated in a case of cultured rat pheochromocytoma (PC12) cells. SiO2 particles are gradually covered with specific protein, fibronectin (FN), by using electrostatic force with pH of FN solution kept at 4.0-8.5. It is also confirmed that FN coverage is proportional to reaction time. Dip-coating to pre-patterned glass plates, on which straight lines of hydrophobic molecules are arranged, allows the FN-coated SiO2 particles to be autonomously structured in a line-and-space pattern. The width and thickness of assembled particles are 35 µm and a few micrometers, respectively. It is also demonstrated FN is successfully supported to the surface of SiO2 particles throughout the structuring process. PC12 cells are incubated on the glass plate at 100% humidity and 310 K. The structured particles work as a scaffold for PC12 cells. Most cells preferentially adhere to FN-coated SiO2 particles. The adhered cells, which are located at the center of structure, are aligned at regular intervals. Meanwhile, in the case of flat glass region and structured bare particles, the number of adhere cells are remarkably small as compared to that of FN-coated SiO2 particles. It is found that an increase of FN coverage raises the number of adhered cells on the particles. These experimental results demonstrate that the structure of protein-coated particles have a function to induce self-alignment of cells. This indicates subsequent differentiation enables a desired network of neuron on the structure.
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