Neuronal differentiation and long-term culture of the human neuroblastoma line SH-SY5Y

Constantinescu, Radu; Constantinescu, Alexandru-Tudor; Reichmann, Heinz; Janetzky, Bernd

doi:10.1007/978-3-211-73574-9_3

Cited by 132 publications

(148 citation statements)

References 37 publications

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“…Confirming previous data, 28,35 neurotransmitter expression changes were not evident after 5 days of culture ( Figure 5B) but required 10 days to become statistically significant ( Figure 5D), particularly those of neurogenin and tyrosine hydroxylase which suggest a shift toward a dopaminergic phenotype. 36 On the other hand, at the same time point, we no longer observed significant differences in expression of cytoskeletal molecules ( Figure 5E), suggesting that modifications of the cytoskeleton are first required in order to achieve a complete differentiation.…”

Section: Expression Studies On Shsy-5y Cellssupporting

confidence: 90%

“…28 In the current study, the cell line grown on flat silicon (Si-Ctrl) and on nanopatterned surfaces demonstrated significantly lower lactate dehydrogenase production than cells grown on plastic ( Figure 3A) and showed increased metabolic activity which, however, did not reach the level of cells grown on plastic and resembled more the level of those exposed to RA ( Figure 3B). …”

Section: Evaluation Of Nanosurface Biocompatibilitymentioning

confidence: 53%

“…As shown by Constantinescu et al, 28 once differentiated, SHSY-5Y cells express cytoskeletal and neurotransmitter markers typical of dopaminergic neurons. Accordingly, after 5 days of culture on rough nanosurfaces, MAP2 expression was higher compared to other culture conditions and similar to RA-treated cells ( Figure 5A).…”

Section: Expression Studies On Shsy-5y Cellsmentioning

confidence: 80%

“…For the differentiation experiments, the concentration of FBS was 10% in accordance with Constantinescu et al 28 For primary podocyte cultures, glomeruli from healthy 2-month-old male Sprague Dawley rats and C57BL6 mice were isolated by sieving, seeded in culture flasks precoated with collagen type IV, and covered by Dulbecco's Modified Eagle's Medium/F12 medium (DMEM/F12) supplemented with 10% FBS, 5 µg/mL transferrin, 10 -7 M hydrocortisone, 5 ng/mL sodium selenite, 0.12 U/mL insulin, 100 µg/mL penicillin, 100 µg/mL streptomycin, and 2 mM l-glutamine (all from Sigma-Aldrich Co.). After 10 days, first-passage podocytes were separated from glomeruli by an additional sieving through a 40 µm mesh (BD Falcon; BD Biosciences; Discovery Labware, Bedford, MA, USA) and seeded onto the nanosurfaces or on plastic.…”

Section: Characteristics Of Nanopatterned Surfacesmentioning

confidence: 99%

“…One microgram of total RNA was reverse transcribed using MMLV reverse transcriptase (Thermo Fisher Scientific) and utilized for semiquantitative real-time polymerase chain reaction, as described. 33 Human primers were the same as described by Constantinescu et al 28 Human GAPDH was utilized as house-keeping gene with the following primers: 5′-CCC ATCACCATCTTCCAGGAG-3′ (forward) and 5′-CTTCT CCATGGTGGTGAAGACG-3′ (reverse) (NM_002046.5).…”

Section: Gene Expression Studymentioning

confidence: 99%

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A nanoporous surface is essential for glomerular podocyte differentiation in three-dimensional culture

Zennaro¹,

Rastaldi²,

Bakeine³

et al. 2016

IJN

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Although it is well recognized that cell–matrix interactions are based on both molecular and geometrical characteristics, the relationship between specific cell types and the three-dimensional morphology of the surface to which they are attached is poorly understood. This is particularly true for glomerular podocytes – the gatekeepers of glomerular filtration – which completely enwrap the glomerular basement membrane with their primary and secondary ramifications. Nanotechnologies produce biocompatible materials which offer the possibility to build substrates which differ only by topology in order to mimic the spatial organization of diverse basement membranes. With this in mind, we produced and utilized rough and porous surfaces obtained from silicon to analyze the behavior of two diverse ramified cells: glomerular podocytes and a neuronal cell line used as a control. Proper differentiation and development of ramifications of both cell types was largely influenced by topographical characteristics. Confirming previous data, the neuronal cell line acquired features of maturation on rough nanosurfaces. In contrast, podocytes developed and matured preferentially on nanoporous surfaces provided with grooves, as shown by the organization of the actin cytoskeleton stress fibers and the proper development of vinculin-positive focal adhesions. On the basis of these findings, we suggest that in vitro studies regarding podocyte attachment to the glomerular basement membrane should take into account the geometrical properties of the surface on which the tests are conducted because physiological cellular activity depends on the three-dimensional microenvironment.

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Section: Expression Studies On Shsy-5y Cellssupporting

confidence: 90%

Section: Evaluation Of Nanosurface Biocompatibilitymentioning

confidence: 53%

Section: Expression Studies On Shsy-5y Cellsmentioning

confidence: 80%

Section: Characteristics Of Nanopatterned Surfacesmentioning

confidence: 99%

Section: Gene Expression Studymentioning

confidence: 99%

See 3 more Smart Citations

A nanoporous surface is essential for glomerular podocyte differentiation in three-dimensional culture

Zennaro¹,

Rastaldi²,

Bakeine³

et al. 2016

IJN

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Biomimetic Scaffolds for Spinal Cord Applications Exhibit Stiffness‐Dependent Immunomodulatory and Neurotrophic Characteristics

Woods

O'Connor

Frugoli

et al. 2021

Adv Healthcare Materials

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After spinal cord injury (SCI), tissue engineering scaffolds offer a potential bridge for regeneration across the lesion and support repair through proregenerative signaling. Ideal biomaterial scaffolds that mimic the physicochemical properties of native tissue have the potential to provide innate trophic signaling while also minimizing damaging inflammation. To address this challenge, taking cues from the spinal cord's structure, the proregenerative signaling capabilities of native cord components are compared in vitro. A synergistic mix of collagen‐IV and fibronectin (Coll‐IV/Fn) is found to optimally enhance axonal extension from neuronal cell lines (SHSY‐5Y and NSC‐34) and induce morphological features typical of quiescent astrocytes. This optimal composition is incorporated into hyaluronic acid scaffolds with aligned pore architectures but varying stiffnesses (0.8–3 kPa). Scaffolds with biomimetic mechanical properties (<1 kPa), functionalized with Coll‐IV/Fn, not only modulate primary astrocyte behavior but also stimulate the production of anti‐inflammatory cytokine IL‐10 in a stiffness‐dependent manner. Seeded SHSY‐5Y neurons generate distributed neuronal networks, while softer biomimetic scaffolds promote axonal outgrowth in an ex vivo model of axonal regrowth. These results indicate that the interaction of stiffness and biomaterial composition plays an essential role in vitro in generating repair‐critical cellular responses and demonstrates the potential of biomimetic scaffold design.

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Eumelanin‐Coated Aligned PLA Electrospun Microfibers to Guide SH‐SY5YCells Spreading, Alignment, And Maturation

Mariano

Fasolino

Dinger

et al. 2023

Adv Materials Inter

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During the development of the nervous system, neurons sense and respond to topographical and biochemical cues found in the brain's native environment. Such niche‐specific cues are pivotal for neural tissue formation and development and have been now widely exploited in neural tissue engineering to develop biologically‐inspired scaffolds able to trigger a desired neuronal behavior. In particular, anisotropic aligned fibers, recapitulating the design of extending axonal tracts or aligned fibers found in the extracellular environment, have emerged as ideal candidates to guide cell alignment and elongation along the substrate main axis as well as promote neuronal differentiation. Among natural polymers used for coatings, melanins, including eumelanin, have been shown to possess antioxidant, anti‐inflammatory, immunomodulatory, and photo‐protective properties. Here, aligned poly(lactic acid) (PLA) fibers are fabricated via electrospinning and then coated with eumelanin via spin coating. Eumelanin‐coated aligned PLA fibers are highly biocompatible and greatly influenced the adhesion, morphology, and spreading of SH‐SY5Y neuroblastoma cells. Furthermore, the eumelanin coating is crucial in promoting cell alignment at the cell‐material interface and SH‐SY5Y cell maturation towards a more mature neuronal phenotype.

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Neuronal differentiation and long-term culture of the human neuroblastoma line SH-SY5Y

Cited by 132 publications

References 37 publications

A nanoporous surface is essential for glomerular podocyte differentiation in three-dimensional culture

A nanoporous surface is essential for glomerular podocyte differentiation in three-dimensional culture

Biomimetic Scaffolds for Spinal Cord Applications Exhibit Stiffness‐Dependent Immunomodulatory and Neurotrophic Characteristics

Eumelanin‐Coated Aligned PLA Electrospun Microfibers to Guide SH‐SY5YCells Spreading, Alignment, And Maturation

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