Differentiation of SH-SY5Y neuronal cells with All-trans-retinoic acid- First Report of 90 days survival of differentiated neuronal cells

Published: 12-06-2020| Version 1 | DOI: 10.17632/spyw8ppd6z.1
Soha Ahmadi,
Qusai Hassan,
Kagan Kerman


A reliable in vitro system that mimics the neuronal function in the brain is a vital aspect of neurobiology research, and it is the key to successful drug discovery. Neuroblastoma SH-SY5Y cells are commonly used to provide an in vitro model of the neurons.[1,2] SH-SY5Y cells can be differentiated to neurons using different chemical and biochemical reagents.[1–5] Using all-trans-retinoic acid (ATRA), a metabolite of vitamin A, to promote the differentiation of neuroblastoma cells to neurons is a simple and cost-effective way but can not produce mature neurons and the differentiated neurons only survive for few days.[4-7] We developed a method that SH-SY5Y cells gradually differentiated to mature neurons using ATRA. The differentiated SH-SY5Y cells grew on a thin layer of extracellular matrix (ECM), which was secreted into the matrix by the cells. The ECM scaffold allowed the neurons to grow in all dimensions with long neurites, which mimicked the brain network. The neuronal 3D in vitro system was maintained for more than 90 days. To our best of knowledge, this is the first report of survival of the mature neurons for 90 days without passaging. The morphology of SH-SY5Y cells during the differentiation process was monitored by phase-contrast microscopy images. The data provided here contained the detail of the method with the microscopy images that were taken during 100 days of the procedure. References: 1. Forster, J. I. et al. Characterization of differentiated SH-SY5Y as neuronal screening model reveals increased oxidative vulnerability. J. Biomol. Screen. 21, 496–509 (2016). 2. Lopes, F. M. et al. Comparison between proliferative and neuron-like SH-SY5Y cells as an in vitro model for Parkinson disease studies. Brain Res. 1337, 85–94 (2010). 3. Presgraves, S. P., Ahmed, T., Borwege, S. & Joyce, J. N. Terminally differentiated SH-SY5Y cells provide a model system for studying neuroprotective effects of dopamine agonists. Neurotox. Res. 5, 579–598 (2003). 4. Korecka, J. A. et al. Phenotypic Characterization of Retinoic Acid Differentiated SH-SY5Y Cells by Transcriptional Profiling. PLoS One 8, e63862 (2013). 5. Shipley, M. M., Mangold, C. A. & Szpara, M. L. Differentiation of the SH-SY5Y human neuroblastoma cell line. J. Vis. Exp. 108, e53193 (2016). 6. Påhlman, S., Ruusala, A. I., Abrahamsson, L., Mattsson, M. E. K. & Esscher, T. Retinoic acid-induced differentiation of cultured human neuroblastoma cells: a comparison with phorbolester-induced differentiation. Cell Differ. 14, 135–144 (1984). 7. Rohwedel, J., Guan, K. & Wobus, A. M. Induction of cellular differentiation by retinoic acid in vitro. Cells Tissues Organs 165, 190–202 (1999).