An Efficient Method for Neuron‐Like Differentiation of
SH
‐
SY5Y
Neuroblastoma Cells Using Retinoic Acid and Laminin‐Rich Extracellular Matrix
Marina Mantellatto Grigoli, Bianca Cruz Pachane, Angelina Maria Fuzer, Sabrina Dorta de Oliveira, Ana Beatriz Aparecida Targas, Vanessa Alexandre‐Silva, Heloisa Sobreiro Selistre‐de‐Araujo, Patricia Regina Manzine, Marcia Regina Cominetti 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.