Differentiated B104 neuroblastoma cells are a high-resolution assay for micropatterned substrates

Abstract

The B104 neuroblastoma cell line was investigated for use as an assay for predicting the patterning of primary neurons. B104 cells were grown on four uniform substrates with the result that the cells preferred, in descending order, poly-D-lysine (PDL), phenyltrichlorosilane (PTCS), coverslip glass, and silicon dioxide coated coverslips. B104 cells were then grown on micropatterned PDL grids on silicon dioxide coated substrates with excellent patterning. Compliance of somata to the pattern, defined as the percentage of cell bodies in a grid field located on the grid pattern, was 86% after 8 h. Neurites were not as compliant, since only 10% of background areas were free of neurites and connected cells. Compliance at longer time periods was greatly reduced. With the addition of the differentiating agent dibutyrylcyclicAMP (DBcAMP), the compliance of somata was maintained at high levels for up to 72 h. Also, the compliance of neurites greatly increased (70%) and showed positive improvement with longer pattern path lengths, contrary to B104 cells without DBcAMP. At longer times neurite compliance was reduced (12% at 28 h and 44% at 72 h). Although there are differences in substrate preferences, the B104 system with DBcAMP appears to be a useful tool in the investigation of the technology of patterned substrates.