Enhancing Neurogenesis of Neural Stem Cells using Homogeneous Nanohole Pattern-modified Conductive Platform

Biocompatible platforms, wherein cells attach and grow, are important for controlling cytoskeletal dynamics and steering stem cell functions, including differentiation. Among various components, membrane integrins play a key role in focal adhesion of cells (18–20 nm in size) and are, thus, highly sensitive to the nanotopographical features of underlying substrates. Hence, it is necessary to develop a platform/technique that can provide high flexibility in controlling nanostructure sizes. We report a platform modified with homogeneous nanohole patterns, effective in guiding neurogenesis of mouse neural stem cells (mNSCs). Sizes of nanoholes were easily generated and varied using laser interference lithography (LIL) by changing the incident angles of light interference on substrates. Among three different nanohole patterns fabricated on conducive transparent electrodes, 500 nm-sized nanoholes showed the best performance for cell adhesion and spreading based on F-actin and lamellipodia/filophodia expression. Enhanced biocompatibility and cell adhesion of these nanohole patterns ultimately resulted in enhancement of neurogenesis of mNSCs, based on the mRNAs expression level of mNSCs marker and several neuronal markers. Therefore, platforms modified with homogeneous nanohole patterns fabricated by LIL are promising for precise tuning of nanostructures of tissue culture platforms and useful for controlling various differentiation lineages of stem cells.