Neuronal networks in vitro: formation and organization on biofunctionalized surfaces

Receptor-mediated recognition of substrate molecules is a prerequisite for nerve cells in order to find their target structures in vivo and leads to f ormation of neuronal connections and networks. In order to study these mech anisms under in vitro conditions, we cultured embryonic hippocampal neurons or neuronal cell lines, SH-SY5Y and PCC7-PCC7-Mz1, onto biofunctionalized surfaces. Micropatterning on polymer surfaces, glass- and silicone-oxide-ba sed chip materials was performed in a micrometer range by microcontact prin ting using polydimethylsiloxane (PDMS) stamps. Hippocampal neurons were fou nd to form networks on chip surfaces under serum-free conditions and remain ed functional for more than a week. Human neuroblastoma cells SH-SY5Y as we ll as PCC7-Mz1 stem cells were found to follow microcontact printed pattern on polystyrene surfaces. Both cell lines showed neuronal marker expression and were cultured for up to 7 days with serum containing culture medium. W idths of 3-5 mu m of coating lines were found to enhance single cell spread ing along the pattern. The techniques described in this study may be useful in promoting nerve cell regeneration and organization following transectio n due to trauma or surgery. The neuronal alignment and network formation in vitro may furthermore serve as a model system in the field of biosensors. (C) 1999 Kluwer Academic Publishers.