Quantitative characterization of phenotypical markers after differentiation of SH-SY5Y cells.

Ducray, Angélique D.; Wiedmer, Linda; Herren, Fabienne; Widmer, Hans Rudolf; Mevissen, Meike (2020). Quantitative characterization of phenotypical markers after differentiation of SH-SY5Y cells. (In Press). CNS & neurological disorders, drug targets Bentham Science Publishers 10.2174/1871527319666200708132716

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BACKGROUND

The human neuroblastoma cell line, SH-SY5Y has been widely used in neuroscience research, especially in studies related to Parkinson's disease. However, differences between clones have been demonstrated, highlighting the importance to characterize the properties of this cell line carefully.

OBJECTIVE

The aim of this study was to characterize the phenotype of undifferentiated and differentiated SH-SY5Y cells using various differentiation protocols.

METHODS

A morphological and a quantitative analysis of markers related to dopaminergic and cholinergic neurons, but also other phenotypes, was performed.

RESULTS

Differentiated cells showed the typical neuronal morphology. Undifferentiated cells expressed low levels of tyrosine hydroxylase (TH) and higher levels of the high-affinity choline transporter (CHT1). Staurosporine (ST)-differentiation resulted in the highest number of TH-immunoreactive cells, followed by phorbol ester phorbol-12-myristate-13-acetat (PMA), whereas differentiation with brain-derived neurotropic factor (BDNF) did not increase TH-immunoreactive cells. TH, dopamine -hydroxylase and vesicular monoamine transporter-2 were also significantly upregulated in ST-differentiated cells compared to both undifferentiated and retinoic acid (RA)-differentiated cells. RA induced the highest number of CHT1-immunoreactive cells while ST- and BDNF-differentiation reduced CHT1-immunoreactive cells, indicating a decrease in the cholinergic phenotype. The presynaptic neuronal protein, αsynuclein, was significantly upregulated in RA- and ST-treated cells compared to undifferentiated cells. Ascorbic acid increased the number of CHT1-immunoreactive cells in all differentiation procedures and ST-differentiated TH-positive cells significantly.

CONCLUSIONS

Our findings indicate that a quantitative characterization of the phenotype is crucial when using SH-SY5Y cells to study the pathogenesis or evaluate compounds for treatment of neurodegenerative diseases.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH) > Veterinary Pharmacology and Toxicology
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Forschungsbereich Mu50 > Forschungsgruppe Neurochirurgie
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH)

UniBE Contributor:

Widmer, Hans Rudolf and Mevissen, Meike

Subjects:

600 Technology > 630 Agriculture
500 Science
500 Science > 570 Life sciences; biology

ISSN:

1996-3181

Publisher:

Bentham Science Publishers

Language:

English

Submitter:

Meike Mevissen

Date Deposited:

01 Sep 2020 14:40

Last Modified:

01 Sep 2020 14:52

Publisher DOI:

10.2174/1871527319666200708132716

PubMed ID:

32640966

Uncontrolled Keywords:

Neuronal differentiation cholinergic neurons dopaminergic neurons parkinson's disease phenotype

BORIS DOI:

10.7892/boris.146247

URI:

https://boris.unibe.ch/id/eprint/146247

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