Differentiation of the SH-SY5Y Human Neuroblastoma Cell Line

Shipley, Mackenzie M.; Mangold, Colleen A.; Szpara, Moriah L.

doi:10.3791/53193-v

Cited by 40 publications

(67 citation statements)

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“…By following the modifications in cell proliferation rate and morphology, we found that SH-SY5Y is fully differentiated 11 days following the beginning of the differentiation protocol, showing a decrease in the proliferation rate and a visible network of branched neurite projections. However, despite the use of B27 supplement and neurobasal medium, many cells do not survive this process, which has been considered a stressful event [ 4 ]. Cells can trigger different responses to stress conditions, including the enhancement of pro-survival pathways and the activation of cell death to eventually remove injured cells [ 33 ].…”

Section: Discussionmentioning

confidence: 99%

“…Human SH-SY5Y neuroblast-like cells are broadly used for in vitro investigation in the field of neurobiology research to overcome all the limitations associated with the culture of primary neurons from the embryonic brain [ 2 , 3 ]. SH-SY5Y cells can be differentiated from a neuroblast-like state (undifferentiated cells) into a neuron-like phenotype (differentiated cells) by using different protocols, including supplementation of retinoic acid (RA) and specific neurotrophins, such as brain-derived neurotrophic factor (BDNF) [ 4 ]. Interestingly, the use of precise protocols for the differentiation of SH-SY5Y cells can select specific neuronal subtypes, including dopaminergic, cholinergic, and adrenergic neurons [ 5 ].…”

Section: Introductionmentioning

confidence: 99%

“…Although SH-SY5Y cells, both undifferentiated and differentiated, are used almost indiscriminately for in vitro experiments that require neuron-like cells, several investigations have described significant differences between the two phenotypes [ 4 ]. Undifferentiated SH-SY5Y cells display a non-polarized morphology with very few short processes.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Manipulation of HSP70-SOD1 Expression Modulates SH-SY5Y Differentiation and Susceptibility to Oxidative Stress-Dependent Cell Damage: Involvement in Oxotremorine-M-Mediated Neuroprotective Effects

et al. 2023

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The differentiation of neural progenitors is a complex process that integrates different signals to drive transcriptional changes, which mediate metabolic, electrophysiological, and morphological cellular specializations. Understanding these adjustments is essential within the framework of stem cell and cancer research and therapy. Human neuroblastoma SH-SY5Y cells, widely used in neurobiology research, can be differentiated into neuronal-like cells through serum deprivation and retinoic acid (RA) supplementation. In our study, we observed that the differentiation process triggers the expression of Heat Shock Protein 70 (HSP70). Notably, inhibition of HSP70 expression by KNK437 causes a dramatic increase in cell death. While undifferentiated SH-SY5Y cells show a dose-dependent decrease in cell survival following exposure to hydrogen peroxide (H2O2), differentiated cells become resistant to H2O2-induced cell death. Interestingly, the differentiation process enhances the expression of SOD1 protein, and inhibition of HSP70 expression counteracts this effect and increases the susceptibility of differentiated cells to H2O2-induced cell death, suggesting that the cascade HSP70-SOD1 is involved in promoting survival against oxidative stress-dependent damage. Treatment of differentiated SH-SY5Y cells with Oxotremorine-M (Oxo), a muscarinic acetylcholine receptor agonist, enhances the expression of HSP70 and SOD1 and counteracts tert–Butyl hydroperoxide-induced cell death and reactive oxygen species (ROS) generation. It is worth noting that co-treatment with KNK437 reduces SOD1 expression and Oxo-induced protection against oxidative stress damage, suggesting the involvement of HSP70/SOD1 signaling in this beneficial effect. In conclusion, our findings demonstrate that manipulation of the HSP70 signal modulates SH-SY5Y differentiation and susceptibility to oxidative stress-dependent cell death and unravels novel mechanisms involved in Oxo neuroprotective functions. Altogether these data provide novel insights into the mechanisms underlying neuronal differentiation and preservation under stress conditions.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Manipulation of HSP70-SOD1 Expression Modulates SH-SY5Y Differentiation and Susceptibility to Oxidative Stress-Dependent Cell Damage: Involvement in Oxotremorine-M-Mediated Neuroprotective Effects

et al. 2023

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“…It has been used as a model cell line for studying neurodegeneration as the cells can be transformed into functional mature neurons. 21 Upon differentiation, the SH-SY5Y cells provide a homogeneous population of mature neurons, which can be used further for various applications. In our experiments, neuronal differentiation of the SH-SY5Y cells was accomplished with retinoic acid treatment, gradual serum deprivation, and the addition of neurotic factors (see Materials and methods section for details).…”

Section: Resultsmentioning

confidence: 99%

Red emitting fluorescent carbon nanoparticles to track spatio-temporal dynamics of endocytic pathways in model neuroblastoma neurons

et al. 2023

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“…SH-SY5Y cells have been extensively used, with numerous studies showing that these cells can be differentiated into specific neuronal subtypes. [69,70] We chose to use this cell line as a robust model for screening our scaffolds for their ability to support neuronal growth.…”

Section: D Neuronal Cell Culture In Scaffoldsmentioning

confidence: 99%

Conducting Polymer‐ECM Scaffolds for Human Neuronal Cell Differentiation

Barberio¹,

Sáez

Withers³

et al. 2022

Adv Healthcare Materials

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3D cell culture formats more closely resemble tissue architecture complexity than 2D systems, which are lacking most of the cell-cell and cell-microenvironment interactions of the in vivo milieu. Scaffold-based systems integrating natural biomaterials are extensively employed in tissue engineering to improve cell survival and outgrowth, by providing the chemical and physical cues of the natural extracellular matrix (ECM). Using the freeze-drying technique, porous 3D composite scaffolds consisting of poly(3,4-ethylene-dioxythiophene) doped with polystyrene sulfonate (PEDOT:PSS), containing ECM components (i.e., collagen, hyaluronic acid, and laminin) are engineered for hosting neuronal cells. The resulting scaffolds exhibit a highly porous microstructure and good conductivity, determined by scanning electron microscopy and electrochemical impedance spectroscopy, respectively. These supports boast excellent mechanical stability and water uptake capacity, making them ideal candidates for cell infiltration. SH-SY5Y human neuroblastoma cells show enhanced cell survival and proliferation in the presence of ECM compared to PEDOT:PSS alone. Whole-cell patch-clamp recordings acquired from differentiated SHSY5Y cells in the scaffolds demonstrate that ECM constituents promote neuronal differentiation in situ. These findings reinforce the usability of 3D conducting supports as engineered highly biomimetic and functional in vitro tissue-like platforms for drug or disease modeling.

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Differentiation of the SH-SY5Y Human Neuroblastoma Cell Line

Cited by 40 publications

References 0 publications

Manipulation of HSP70-SOD1 Expression Modulates SH-SY5Y Differentiation and Susceptibility to Oxidative Stress-Dependent Cell Damage: Involvement in Oxotremorine-M-Mediated Neuroprotective Effects

Manipulation of HSP70-SOD1 Expression Modulates SH-SY5Y Differentiation and Susceptibility to Oxidative Stress-Dependent Cell Damage: Involvement in Oxotremorine-M-Mediated Neuroprotective Effects

Red emitting fluorescent carbon nanoparticles to track spatio-temporal dynamics of endocytic pathways in model neuroblastoma neurons

Conducting Polymer‐ECM Scaffolds for Human Neuronal Cell Differentiation

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