Micropatterning B104 neuroblastoma cells in culture: an assay for neuronal pattern synthesis

Our goal is the reliable synthesis of patterned living neural networks. To assess rapidly the qualities of candidate substrates we use the B104 neuroblastoma cell line with and without the differentiating agent dibutyrylcyclicAMP (DBcAMP). B104 cells prefer, in order, poly-D-lysine (PDL), to phenyltrichlorosilane (PTCS), coverslip glass, and silicon dioxide coated coverslips. At 8 hours micropatterned B104 cells had excellent compliance of somata (86%), but not of neurites (10% of background areas free of neurites). Compliance was greatly reduced thereafter. Addition of DBcAMP increased the compliance of neurites (70%), with patterns maintained up to 72 hours. The assay identified chondroitin sulfate (CSPG) as more cytophobic than mouse fibrinogen, bovine serum albumin (BSA), and glycine, suggesting that a pattern of PDL against CSPG in the background would be optimal. The results are being applied to a culture system of hippocampal neurons