The Effects of Serum Removal on Gene Expression and Morphological Plasticity Markers in Differentiated SH-SY5Y Cells
| Autor: | Alexander T. Sack, Teresa Schuhmann, Alix C. Thomson, Gunter Kenis, Bart P. F. Rutten, Tom A. de Graaf |
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| Přispěvatelé: | RS: FPN CN 4, Cognition, RS: MHeNs - R3 - Neuroscience, Psychiatrie & Neuropsychologie, MUMC+: MA Psychiatrie (3), MUMC+: MA Niet Med Staf Psychiatrie (9) |
| Jazyk: | angličtina |
| Rok vydání: | 2022 |
| Předmět: |
0301 basic medicine
SH-SY5Y Neurite Plasticity Synaptogenesis Gene Expression Biology SH-SY5Y cells 03 medical and health sciences Cellular and Molecular Neuroscience Neuroblastoma 0302 clinical medicine Cell Line Tumor Gene expression Humans Gene Neurons Cell growth Cell Differentiation Serum deprivation Cell Biology General Medicine In vitro Cell biology Human neuron model 030104 developmental biology Signal transduction 030217 neurology & neurosurgery Biomarkers |
| Zdroj: | Cellular and Molecular Neurobiology, 42(6), 1829-1839. Springer Verlag |
| ISSN: | 0272-4340 |
| Popis: | Despite the widespread use of the SH-SY5Y human neuroblastoma cell line in modeling human neurons in vitro, protocols for growth, differentiation and experimentation differ considerably across the literature. Many studies fully differentiate SH-SY5Y cells before experimentation, to investigate plasticity measures in a mature, human neuronal-like cell model. Prior to experimentation, serum is often removed from cell culture media, to arrest the cell growth cycle and synchronize cells. However, the exact effect of this serum removal before experimentation on mature, differentiated SH-SY5Y cells has not yet been described. In studies using differentiated SH-SY5Y cells, any effect of serum removal on plasticity markers may influence results. The aim of the current study was to systematically characterize, in differentiated, neuronal-like SH-SY5Y cells, the potentially confounding effects of complete serum removal in terms of morphological and gene expression markers of plasticity. We measured changes in commonly used morphological markers and in genes related to neuroplasticity and synaptogenesis, particularly in the BDNF-TrkB signaling pathway. We found that complete serum removal from already differentiated SH-SY5Y cells increases neurite length, neurite branching, and the proportion of cells with a primary neurite, as well as proportion of βIII-Tubulin and MAP2 expressing cells. Gene expression results also indicate increased expression of PSD95 and NTRK2 expression 24 h after serum removal. We conclude that serum deprivation in differentiated SH-SY5Y cells affects morphology and gene expression and can potentially confound plasticity-related outcome measures, having significant implications for experimental design in studies using differentiated SH-SY5Y cells as a model of human neurons. |
| Databáze: | OpenAIRE |
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