Substrate stiffness effects on SH‐SY5Y: The dichotomy of morphology and neuronal behavior

Abstract

Like many other cell types, neuroblastoma cells are also known to respond to mechanical cues in their microenvironment in vitro. They were shown to have mechanotransduction pathways, which result in enhanced neuronal morphology on stiff substrates. However, in previous studies, the differentiation process was monitored only by morphological parameters. Motivated by the lack of comprehensive studies that investigate the effects of mechanical cues on neuroblastoma differentiation, we used SH-SY5Y cells differentiated on polyacrylamide (PA) gels as a model. Cells differentiated on the surface of PA hydrogels with three different elastic moduli (0.1, 1, and 50 kPa) were morphologically evaluated and their electrophysiological responsiveness was probed using calcium imaging. Immunodetection of neural marker TUJ1 and p-FAK was used for biochemical characterization. Groups with defined stiffness that are matching and nonmatching to neural tissue extracellular matrix were used to distinguish biomimetic results from other effects. Results show that while cells display morphologies that do not resemble neurons on soft substrates, they are in fact electrophysiologically more responsive and abundant in neuronal marker TUJ1. Our findings suggest that while neuronal differentiation occurs more efficiently in microenvironments mechanically mimicking neural tissue, the SH-SY5Y model demonstrates morphologies that conflict with neuronal behavior under these conditions. These results are expected to contribute considerable input to researchers that use SH-SY5Y as a neuron model.

DDR2 signaling and mechanosensing orchestrate neuroblastoma cell fate through different transcriptome mechanisms

“…We found no changes in gene expression between SH-SY5Y cells grown on a soft 2 kPa versus a hard 20 kPa substrate. This result is rather unexpected given that our results, along with a few previous studies on neuroblastomas cells, have observed overt changes in cell proliferation, cell size, and cell differentiation in response to stiffness changes [47][48][49]. On the other hand, knocking down the collagen receptor DDR2 profoundly altered SH-SY5Y cell transcriptome.…”

Astrocyte 3D culture and bioprinting using peptide functionalized hyaluronan hydrogels

“…Furthermore, while a decrease in metabolic activity was observed between day 1 and day 6, the micrographs show an increase in cell number, indicating proliferation both without and with peptides. The cell clusters appeared less dense over time with more condensed cells, which has previously been observed for undifferentiated SH-SY5Y cultured on soft substrates, also accompanied by a decrease in cell stiffness indicating lower degrees of actin polymerization [ 28 , 29 ]. The hydrogels thus provide adequate and similar support for cell proliferation irrespectively of peptide functionalization, on par with observations of SH-SY5Y in other soft engineered hydrogels.…”

Chemically Triggered Hydrogel Transformations through Covalent Adaptable Networks and Applications in Cell Culture

“…Thus, SH-SY5Y neuronal cells were utilized in cell attachment studies in 2D cell culture, which are known to respond to mechanical cues in their microenvironment in vitro . We observed that SH-SY5Y cells successfully attached onto the surfaces of both H-2 and H-3 .…”