Vapor-based micro/nano-partitioning of fluoro-functional group immobilization for long-term stable cell patterning

Abstract

A technique for partitioning cell adhesion with long-term stability is an important tool in biology and medical experiments. This study has developed a simple vapor-based immobilization method using a compound bearing a fluoro-functional group on a cell culture surface with micro/nanoscale patterns. The vapor of a silane coupling agent containing the fluoro-functional group was introduced into the micro-/nanochannels of polydimethylsiloxane elastomer placed on the target surface in a vacuum desiccator. The agent was immobilized with a pattern shown only on the areas of the channels on the surface. Although it is difficult to introduce the agent solution into a sub-micrometer-width channel via a conventional liquid-based approach, the vapor can be introduced into nanochannels with an exceptionally narrow width of 800 nm and an exceptionally low height of 700 nm. The fluoro-functional group area was clearly found to repel the adhesion of fluorescent-dye-conjugated fibronectin molecules. Four different types of cells, mouse skeletal myoblast cell line C2C12 cells, human cervix adenocarcinoma cell line HeLa cells, bovine artery endothelial cells HH, and mouse mammary gland epithelial cell line NMuMG cells, were prepared and each type was seeded onto the immobilized culture surfaces of glass and polystyrene dishes. The cells were adhered on the non-fluoro-functional group immobilized area and the cell pattern was maintained during atleast a 39 days of cultivation. This method is expected to be useful for simple and stable cell patterning in biological and medical fields.