USE OF CHEMICALLY PATTERNED SUBSTRATE TO STUDY DIRECTIONAL EFFECT OF DAMAGING ELECTRICAL-STIMULATION ON CULTURED NEUROBLASTOMA-CELLS

We used ordered arrangements of neuroblastoma cells in culture on chemically patterned substrates to direct the orientation of electrical stimulation with respect to cell alignment. Chemically patterned parallel lines of self-assembled monolayer films were fabricated on glass substrates via a deep UV lithographic procedure. Cultured neuroblastoma cells deposited on these substrates formed long(similar to 300 mu m in length) neuritic processes along the patterned lines in the presence of retinoic acid. Cells attached to the surface of these substrates were placed in a stimulation chamber so that an electric field (1.4-1.9 V/cm) could be applied in the direction parallel or perpendicular to neuritic orientation. A majority of cells aligned parallel to the orientation of electrical stimulation exhibited a variety of cellular responses including neuritic tip damage, reductions in neuritic length and varicosity formation. These effects were observed to a lesser degree on the cells when electrical stimulation with the same magnitude was applied perpendicularly to the cell alignment. This work supports earlier findings that geometry is a crucial factor in determining cellular response to applied electric fields and goes on to show that cellular orientation is a key factor in determining cellular damage in culture.