Brain-derived neurotrophic factor (BDNF) promotes molecular polarization and differentiation of immature neuroblastoma cells into definitive neurons

Hromádková, Lenka; Bezděková, Dagmar; Pala, Jan; Schedin‐Weiss, Sophia; Tjernberg, Lars O.; Höschl, Cyril; Ovsepian, Saak V.

doi:10.1016/j.bbamcr.2020.118737

Cited by 43 publications

(32 citation statements)

References 54 publications

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“…We choose for our studies the SH-SY5Y neuroblastoma cell line since it is a well-established cellular model for experimental neurobiology studies, including metabolism, neural differentiation and neurodegenerative processes (Encinas et al, 2002). In fact, the sequential treatment of these cells with retinoic acid (RA) and human brain−derived neurotrophic factor (hBDNF), causes differentiation in a neuron-like phenotype with neuritic processes (Hromadkova et al, 2020). High quality fullcovered WS 2 and graphene samples were grown on sapphire via CVD, and bare sapphire substrates and culture wells were used as controls to test the material effect on cell viability and differentiation.…”

Section: Introductionmentioning

confidence: 99%

Effect of Chemical Vapor Deposition WS2 on Viability and Differentiation of SH-SY5Y Cells

et al. 2020

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In recent years, transition metal dichalcogenides have been attracting an increasing interest in the biomedical field, thus implying the need of a deeper understanding of their impact on cell behavior. In this study we investigate tungsten disulfide (WS 2) grown via chemical vapor deposition (CVD) on a transparent substrate (sapphire) as a platform for neural-like cell culture. We culture SH-SY5Y human neuroblastoma cells on WS 2 , using graphene, sapphire and standard culture well as controls. The quality, thickness and homogeneity of the materials is analyzed using atomic force microscopy and Raman spectroscopy. The cytocompatibility of CVD WS 2 is investigated for the first time by cell viability and differentiation assessment on SH-SY5Y cells. We find that cells differentiated on WS 2 , displaying a viability and neurite length comparable with the controls. These findings shine light on the possibility of using WS 2 as a cytocompatible material for interfacing neural cells.

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Section: Introductionmentioning

confidence: 99%

Effect of Chemical Vapor Deposition WS2 on Viability and Differentiation of SH-SY5Y Cells

et al. 2020

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“…Therefore, we treated the HA-MOP SH-SY5Y cell line with all-trans retinoic acid for 5 days, followed by BDNF treatment for 5 days, which differentiated SH-SY5Y cells into a state similar to a primary culture (Figure 5A). 24,25 Upon differentiation, SH-SY5Y cells became enriched with prolonged neurites, whose morphological characteristics resembled those of the primary culture (arrowheads on the rightmost panel in Figure 5A).…”

Section: Continuous Opioid Administration Lowered Mop Expression On T...mentioning

confidence: 86%

CRISPR/Cas9 unveils the dynamics of the endogenous µ‐opioid receptors on neuronal cells under continuous opioid stimulation

Shimizu

Shiraki

2022

Pharmacology Res & Perspec

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Long‐term opioid use develops tolerance and attenuates analgesic effects. Upon activation, µ‐opioid receptors (MOPs) are internalized and directed to either recycling or degradation pathway. Ligand stimulation also promotes de novo MOP synthesis. These processes collaboratively regulate MOP expression and play critical roles in tolerance development. However, there is limited understanding of how the endogenous MOP expression changes after prolonged opioid administration because previous analyses have focused on individual processes using overexpression systems, which ignored physiological regulation. Another fundamental problem is the unavailability of commercial antibodies to detect the low expression of endogenous MOP in neuronal systems. Here, we established a neuronal cell line to detect endogenous MOP with sufficient sensitivity using CRISPR/Cas9 technology. We incorporated the hemagglutinin sequence into the MOP gene of the SH‐SY5Y cell. The genome‐editing did not significantly impair MOP functions such as MOP internalization or the downstream signaling. The clone was differentiated into a state similar to the primary culture undergoing treatment with all‐trans retinoic acid, followed by brain‐derived neurotrophic factor. Upon continuous stimulation with MOP ligands, endogenous MOP constantly decreased up to 48 h. The expression level was maintained at a certain level following this period, depending on the ligand properties. DAMGO reduced MOP from the cell surface by about 70%, while morphine did so by 40%. Our results indicate that even a few days of opioid administration could significantly reduce the MOP expression level. Our cell line could be a potential tool to investigate the molecular mechanisms underlying the problems caused by long‐term opioid use.

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“…To assess the differentiation of SH-SY5Y cells on the GelMA hydrogel, the cells were stained with

-tubulin III and NucGreen®. Despite

-tubulin III being an established neuronal cell marker, extending this assessment by microtubule-associated protein 2 (MAP2) and Tau protein (TAU) staining would provide valuable information regarding the cells’ states of differentiation [ 20 , 21 , 22 , 23 , 24 , 25 ]. To determine the migration, z-stacks were generated from 0 to 60 µm.…”

Section: Discussionmentioning

confidence: 99%

Stiff-to-Soft Transition from Glass to 3D Hydrogel Substrates in Neuronal Cell Culture

Akçay

Lüttge

2021

Micromachines

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Over the past decade, hydrogels have shown great potential for mimicking three- dimensional (3D) brain architectures in vitro due to their biocompatibility, biodegradability, and wide range of tunable mechanical properties. To better comprehend in vitro human brain models and the mechanotransduction processes, we generated a 3D hydrogel model by casting photo-polymerized gelatin methacryloyl (GelMA) in comparison to poly (ethylene glycol) diacrylate (PEGDA) atop of SH-SY5Y neuroblastoma cells seeded with 150,000 cells/cm2 according to our previous experience in a microliter-sized polydimethylsiloxane (PDMS) ring serving for confinement. 3D SH-SY5Y neuroblastoma cells in GelMA demonstrated an elongated, branched, and spreading morphology resembling neurons, while the cell survival in cast PEGDA was not supported. Confocal z-stack microscopy confirmed our hypothesis that stiff-to-soft material transitions promoted neuronal migration into the third dimension. Unfortunately, large cell aggregates were also observed. A subsequent cell seeding density study revealed a seeding cell density above 10,000 cells/cm2 started the formation of cell aggregates, and below 1500 cells/cm2 cells still appeared as single cells on day 6. These results allowed us to conclude that the optimum cell seeding density might be between 1500 and 5000 cells/cm2. This type of hydrogel construct is suitable to design a more advanced layered mechanotransduction model toward 3D microfluidic brain-on-a-chip applications.

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Brain-derived neurotrophic factor (BDNF) promotes molecular polarization and differentiation of immature neuroblastoma cells into definitive neurons

Cited by 43 publications

References 54 publications

Effect of Chemical Vapor Deposition WS2 on Viability and Differentiation of SH-SY5Y Cells

Effect of Chemical Vapor Deposition WS2 on Viability and Differentiation of SH-SY5Y Cells

CRISPR/Cas9 unveils the dynamics of the endogenous µ‐opioid receptors on neuronal cells under continuous opioid stimulation

Stiff-to-Soft Transition from Glass to 3D Hydrogel Substrates in Neuronal Cell Culture

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