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 chem ically patterned substrates to direct the orientation of electrical st imulation with respect to cell alignment. Chemically patterned paralle l lines of self-assembled monolayer films were fabricated on glass sub strates via a deep UV lithographic procedure. Cultured neuroblastoma c ells deposited on these substrates formed long(similar to 300 mu m in length) neuritic processes along the patterned lines in the presence o f retinoic acid. Cells attached to the surface of these substrates wer e 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 ne uritic orientation. A majority of cells aligned parallel to the orient ation of electrical stimulation exhibited a variety of cellular respon ses including neuritic tip damage, reductions in neuritic length and v aricosity formation. These effects were observed to a lesser degree on the cells when electrical stimulation with the same magnitude was app lied perpendicularly to the cell alignment. This work supports earlier findings that geometry is a crucial factor in determining cellular re sponse to applied electric fields and goes on to show that cellular or ientation is a key factor in determining cellular damage in culture.