Containment and growth of neuroblastoma cells on chemically patterned substrates

Patterned substrates offer the promise of controlled positioning and directional guidance of growing neurites. Therefore, they could be useful for constructing small neuronal networks with defined geometry in vitro. We have fabricated chemically patterned substrates using self-assembled monolayer films with a lithographic mask technique and demonstrated the feasibility for geometrically patterning neuroblastoma cells in culture. N-octadecyltrichlorosilane (OTS) was chemically bonded to glass and fused silica substrates, rendering the surface hydrophobic and non-adhesive to cells. Using surface analysis techniques, we have confirmed that OTS films were true monolayers and can be photocleaved from the surface by deep UV irradiation. An adhesive pattern of n-(2-aminoethyl-3-aminopropyl)trimethoxysilane was formed on a selectively irradiated OTS surface via a deep UV lithographic procedure. The chemically patterned surface was then seeded with SK-N-SH human neuroblastoma cells, and cellular attachment and growth were monitored by optical microscopy. The use of 2-dimensional substrates supported the containment and growth of neuroblastoma cells within the pattern for at least 15 days in culture. These chemical patterns may also be useful in controlled arrangements of hearts cells or muscle cells on prosthetic implant devices.