An In Vitro Model for Neuroscience: Differentiation of SH-SY5Y Cells into Cells with Morphological and Biochemical Characteristics of Mature Neurons

Article type: Research Article

Authors: Agholme, Lotta^{a; *} | Lindström, Tobias^{a; b} | Kågedal, Katarina^c | Marcusson, Jan^a | Hallbeck, Martin^{b; d}

Affiliations: [a] Department of Clinical and Experimental Medicine, Division of Geriatrics, Faculty of Health Science, Linköping University, Linköping, Sweden | [b] }Department of Clinical and Experimental Medicine, Division of Clinical Pathology, Faculty of Health Science, Linköping University, Linköping, Sweden | [c] Department of Clinical and Experimental Medicine, Division of Experimental Pathology, Faculty of Health Science, Linköping Univeristy, Linköping, Sweden | [d] Department of Pathology, Linköping University Hospital, Linköping, Sweden

Correspondence: [*] Correspondence to: Lotta Agholme, Department of Clinical and Experimental Medicine, Division of Geriatrics, Faculty of Health science, Linköping University, SE-581 85 Linköping, Sweden. Tel.: +46 10 1031500; Fax: +46 10 1031529; E-mail: [email protected].

Note: [] Handling Associate Editor: Thomas Shea

Abstract: Neuroscience, including research on Alzheimer's disease, is hampered by the lack of suitable in vitro models to study the human nervous system. To counteract this, many attempts to differentiate cell lines into more neuron-like cells have been performed, resulting in partial expression of neuronal features. Furthermore, it has been reported that neuroblastoma cell lines lack mature isoforms of tau. Our aim was to develop an improved in vitro model, generating sustainable cells with morphology and biochemistry of human, mature neurons. To obtain cells with neuronal differentiation and function, we investigated the effect of combining three-dimensional culturing of SH-SY5Y cells in extracellular matrix (ECM) gel with several factors reported to have neuro-differentiating effects. This resulted in cells with apparent neuronal morphology with long, extensively branched neurites. Further investigation revealed expression of several neurospecific markers including synapse protein Sv2 and nuclear marker NeuN, as well as the presence of synapses and axonal vesicle transport. In addition, these cells expressed mature tau isoforms, and tau protein expression was significantly increased compared to undifferentiated cells, reaching levels found in adult human brain. In conclusion, we found that pre-treatment with retinoic acid followed by ECM gel culturing in combination with brain derived neurotrophic factor, neuregulin β1, nerve growth factor, and vitamin D3 treatment generated sustainable cells with unambiguous resemblance to adult neurons. These cells also expresses adult splicing forms of tau with neuronal localization, making this cellular in vitro model useful in many areas of neuroscience research, particularly the Alzheimer's disease field.

Keywords: Alzheimer's disease, differentiation, in vitro model, neuroblastoma, tau

DOI: 10.3233/JAD-2010-091363

Journal: Journal of Alzheimer's Disease, vol. 20, no. 4, pp. 1069-1082, 2010