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Open AccessArticle

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

by and *,†
Neuro-Nanoscale Engineering, Department of Mechanical Engineering and Institute of Complex Molecular Systems (ICMS), Eindhoven University of Technology, 5600 MB Eindhoven, The Netherlands
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Academic Editor: Colin Dalton
Micromachines 2021, 12(2), 165; https://doi.org/10.3390/mi12020165
Received: 3 December 2020 / Revised: 26 January 2021 / Accepted: 5 February 2021 / Published: 8 February 2021
(This article belongs to the Special Issue Microfluidic Brain-on-a-Chip)
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. View Full-Text
Keywords: hydrogel; 3D cell culture; brain-on-a-chip; SH-SY5Y cells; GelMA; PEGDA; mechanotransduction hydrogel; 3D cell culture; brain-on-a-chip; SH-SY5Y cells; GelMA; PEGDA; mechanotransduction
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MDPI and ACS Style

Akcay, G.; Luttge, R. Stiff-to-Soft Transition from Glass to 3D Hydrogel Substrates in Neuronal Cell Culture. Micromachines 2021, 12, 165. https://doi.org/10.3390/mi12020165

AMA Style

Akcay G, Luttge R. Stiff-to-Soft Transition from Glass to 3D Hydrogel Substrates in Neuronal Cell Culture. Micromachines. 2021; 12(2):165. https://doi.org/10.3390/mi12020165

Chicago/Turabian Style

Akcay, Gulden; Luttge, Regina. 2021. "Stiff-to-Soft Transition from Glass to 3D Hydrogel Substrates in Neuronal Cell Culture" Micromachines 12, no. 2: 165. https://doi.org/10.3390/mi12020165

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