Retinoic acid-evoked differentiation of neuroblastoma cells predominates over growth factor stimulation: An automated image capture and quantitation approach to neuritogenesis

To facilitate the characterization of compounds that have positive growth f actor mimetic effects on neuritogenesis, we have implemented a high-through put functional assay which measures, in a multiparametric manner, the proli feration and differentiation characteristics of cells in a microtiter plate . Conditions were established using chronic incubation of SH-SY5Y human neu roblastoma cells with retinoic acid (RA) and/or nerve growth factor (NGF) i n which discernible alterations in proliferation, growth, and differentiati on of cells were induced. SH-SY5Y cells were fixed and labeled by immunocyt ochemistry, and an automated image acquisition and analysis package on Cell omics ArrayScanII was utilized to quantify the effects of these treatments on cell characteristics. NGF and retinoic acid were found to increase multi ple parameters of SH-SY5Y differentiation, including an increased proportio n of cells having neurites and increased extent of branching. However, mark ed differences in the effects of these compounds on SH-SY5Y growth and diff erentiation were also detected: whereas NGF increased cell number, RA treat ment decreased cell number, and RA but not NGF caused significant elongatio n of neurites. This study quantifies and characterizes the effects of diffe rentiating and proliferating agents on a human-derived neuroblastoma cell l ine. The high-content, rapid-throughput nature of this assay makes it ideal for functional identification and characterization of compounds regulating cell behavior. (C) 2001 Academic Press.