Cell patterning via optimized dielectrophoretic force within hexagonal electrodes in vitro for skin tissue engineering

Tissue reconstruction through in vitro cell seeding is a popular method for tissue engineering. In this paper, we proposed a thin-layer structure consisting of multiple hexagons for the regeneration of skin tissue. Cells could be seeded and cultured within the structure via dielectrophoresis (DEP) actively. A thin layer of the structure was fabricated with biocompatible medical-grade stainless steel via precise laser cutting. The fabricated layers were stacked together to form a 3D electrode pair, which could be used to generate a 3D electric field. Thus, the suspended cells within the structure could be patterned via DEP manipulation. The input voltage was examined and optimized to ensure cell viability and patterning efficiency during the DEP manipulation process. As soon as we applied the optimized voltage, human foreskin fibroblast (HFF) cells could be attracted along the edge of the electrodes, forming hexagonal cellular patterns. After that, a single layer of the patterned cells was further cultured in an incubator for 7 days and observed under a microscope. The obtained images showed that the seeded cells could proliferate and fill in the hexagonal wells, which could be used for further skin tissue regeneration. As shown in the experimental results, this structure could be used for active cell seeding and proliferation for the development of skin tissue engineering.