An efficient method for neuron-like differentiation of SH-SY5Y neuroblastoma cells using retinoic acid and laminin-rich extracellular matrix

An efficient method for neuron-like differentiation of SH-SY5Y neuroblastoma cells using retinoic acid and laminin-rich extracellular matrix | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL This is a preprint and has not been peer reviewed. Data may be preliminary. 6 March 2025 V1 Latest version Share on An efficient method for neuron-like differentiation of SH-SY5Y neuroblastoma cells using retinoic acid and laminin-rich extracellular matrix Authors : Marina Grigoli , Bianca Cruz Pachane 0000-0002-7302-4498 , Angelina Fuzer , Sabrina de Oliveira , Ana Beatriz Targas , Vanessa Silva , Heloisa Selistre-de-Araujo , Patricia Manzine , and Marcia Cominetti 0000-0001-6385-7392 [email protected] Authors Info & Affiliations https://doi.org/10.22541/au.174126909.99331296/v1 388 views 125 downloads Contents Abstract Supplementary Material Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract The SH-SY5Y neuroblastoma cell line is a widely used in vitro model for studying neuronal differentiation and neurodegenerative conditions such as Alzheimer’s disease (AD) due to its ability to undergo neuronal-like differentiation and express key neuronal markers relevant to AD pathology. However, current differentiation protocols often lack physiological relevance, primarily due to the absence of extracellular matrix (ECM) components critical for mechanotransduction and cellular adhesion. This study describes a novel and efficient method for differentiating SH-SY5Y cells into neuron-like cells by combining retinoic acid (RA) treatment with a laminin-rich ECM (LrECM). Our approach significantly enhanced neuronal differentiation markers, including β3-tubulin and NeuN, and promoted extensive neurite outgrowth. Immunofluorescence analysis showed increased expression of these markers, confirming enhanced neuronal characteristics. Western blot results further validated sustained β3-tubulin expression over time. Adding LrECM also augmented cholinergic differentiation, as evidenced by increased acetylcholinesterase activity measured through colorimetric assays. The findings highlight the critical role of LrECM in enhancing neuronal differentiation and maintaining neuron-like characteristics over longer periods. This approach provides a valuable improvement for refining in vitro models used in the study of neuronal development and neurodegenerative diseases, particularly AD. Supplementary Material File (grigoli_et_al_ejn.pdf) Download 784.01 KB Information & Authors Information Version history V1 Version 1 06 March 2025 Copyright This work is licensed under a Non Exclusive No Reuse License. Keywords extracellular matrix laminin neuronal differentiation retinoic acid sh-sy5y Authors Affiliations Marina Grigoli Universidade Federal de São Carlos View all articles by this author Bianca Cruz Pachane 0000-0002-7302-4498 Barretos Cancer Hospital View all articles by this author Angelina Fuzer Universidade Federal de São Carlos View all articles by this author Sabrina de Oliveira Universidade Federal de São Carlos View all articles by this author Ana Beatriz Targas Universidade Federal de São Carlos View all articles by this author Vanessa Silva Universidade Federal de São Carlos View all articles by this author Heloisa Selistre-de-Araujo Universidade Federal de São Carlos View all articles by this author Patricia Manzine Universitat de Barcelona Faculty of Pharmacy and Food Science View all articles by this author Marcia Cominetti 0000-0001-6385-7392 [email protected] Universidade Federal de São Carlos View all articles by this author Metrics & Citations Metrics Article Usage 388 views 125 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Marina Grigoli, Bianca Cruz Pachane, Angelina Fuzer, et al. An efficient method for neuron-like differentiation of SH-SY5Y neuroblastoma cells using retinoic acid and laminin-rich extracellular matrix. Authorea . 06 March 2025. DOI: https://doi.org/10.22541/au.174126909.99331296/v1 If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. Simply select your manager software from the list below and click Download. For more information or tips please see 'Downloading to a citation manager' in the Help menu . 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