Open AccessQuantitative Characterization of Phenotypical Markers After Differentiation of SH-SY5Y Cells
Author(s) -
Angélique Ducray,
Linda Wiedmer,
Fabienne L Herren,
Hans Rudolf Widmer,
Meike Mevissen
Publication year - 2020
Publication title -
cns and neurological disorders. drug targets
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 86
eISSN - 1996-3181
pISSN - 1871-5273
DOI - 10.2174/1871527319666200708132716
Subject(s) - cellular differentiation , glial cell line derived neurotrophic factor , tyrosine hydroxylase , biology , retinoic acid , microbiology and biotechnology , cell culture , choline acetyltransferase , endocrinology , phenotype , ascorbic acid , stem cell , medicine , neurotrophic factors , dopamine , cholinergic , biochemistry , receptor , food science , gene , genetics
Background: The human neuroblastoma cell line, SH-SY5Y, has been widely used in neuroscienceresearch, especially in studies related to Parkinson's disease. However, differences betweenclones have been demonstrated, highlighting the importance to characterize the properties of this cellline carefully. Objective: The aim of this study was to characterize the phenotype of undifferentiated and differentiatedSH-SY5Y cells using various differentiation protocols. Methods: A morphological and quantitative analysis of markers related to dopaminergic and cholinergicneurons, but also other phenotypes, was performed. Results: Differentiated cells showed the typical neuronal morphology. Undifferentiated cells expressedlow levels of Tyrosine Hydroxylase (TH) and higher levels of the high-affinity CholineTransporter (CHT1). Staurosporine (ST)-differentiation resulted in the highest number of THimmunoreactivecells, followed by phorbol ester Phorbol-12-Myristate-13-Acetate (PMA), whereasdifferentiation with Brain-Derived Neurotrophic Factor (BDNF) did not increase TH-immunoreactivecells. TH, dopamine β-hydroxylase and vesicular monoamine transporter-2 were also significantly upregulatedin ST-differentiated cells compared to both undifferentiated and Retinoic Acid (RA)-differentiated cells. RA induced the highest number of CHT1-immunoreactive cells while ST- andBDNF-differentiation reduced CHT1-immunoreactive cells, indicating a decrease in the cholinergic phenotype.The presynaptic neuronal protein, α-synuclein, was significantly upregulated in RA- and ST-treatedcells compared to undifferentiated cells. Ascorbic acid increased the number of CHT1-immunoreactivecells in all differentiation procedures and ST-differentiated TH-positive cells significantly. Conclusions: Our findings indicate that a quantitative characterization of the phenotype is crucial whenusing SH-SY5Y cells to study the pathogenesis or evaluate compounds for treatment of neurodegenerativediseases.