Fabrication of device equipped with rope-like axon bundles by diamond-like carbon thin film patterning deposition

The aim of this research is to fabricate plane-type devices possessing straight longer axons individually and regularly arrayed for evaluating the axon behavior of neuronal cells in vitro toward the development of brain-on-chip models, utilizing diamond-like carbon (DLC) thin film patterning deposition. The DLC thin films were deposited on a polydimethylsiloxane (PDMS) plate with a plasma CVD method under four conditions, with/without stretching of and a metal mask on the plate, and the fabricated substrates were used to culture human neuroblastoma cells, SH-SY5Y, for up to 21 days. In the case of the stretched PDMS plate with the mask covered, extremely peculiar structures were created on the DLC deposited areas of the substrate, with multiple 20 μm-thick rope-like axons aligned straight at the length of millimeter-level at intervals of about 200 μm. It was found that the rope-like axons consisted of plenty of thinner axons, and were supported by the clusters of cells at both ends. It was strongly suggested that the linear wrinkle structures created by the DLC deposition played an important role in the formation of the rope-like axons. This device can be fabricated by means of not conventional lithographic techniques, but only DLC thin film deposition.