An Efficient Method for Neuron-Like Differentiation of SH-SY5Y Neuroblastoma Cells Using Retinoic Acid and Laminin-Rich Extracellular Matrix

Abstract

The human neuroblastoma cell line SH- SY5Y is widely used in vitro as a model due to its ability to acquire neuron- like properties. Conventional differentiation protocols often rely on retinoic acid (RA), which can lead to limited stability of neuronal- like phenotypes over time. In this study, we evaluated a differentiation approach combining RA with a laminin- rich extracellular matrix (LrECM). This approach accelerated differentiation, with neuron- like morphology and neurite outgrowth observed as early as 4 days and sustained features for up to 10 days. Immunofluorescence analysis indicated increased NeuN expression, and Western blot confirmed the sustained presence of β3- tubulin during differentiation. Furthermore, a trend toward enhanced acetylcholinesterase (AChE) activity suggested partial cholinergic features. Altogether, these findings indicate that incorporating LrECM supports the stability of RA- induced neuron- like characteristics in SH- SY5Y cells and may provide a useful alternative for studies of neuronal–phenotype differentiation.