Substrate stiffness affects the morphology and gene expression of epidermal neural crest stem cells in a short term culture

Abstract

According to the intrinsic plasticity of stem cells, controlling their fate is a critical issue in cell‐based therapies. Recently, a growing body of evidence has suggested that substrate stiffness can affect the fate decisions of various stem cells. Epidermal neural crest stem cells as one of the main neural crest cell derivatives hold great promise for cell therapies due to presenting a high level of plasticity. This study was conducted to define the influence of substrate stiffness on the lineage commitment of these cells. Here, four different polyacrylamide hydrogels with elastic modulus in the range of 0.7–30 kPa were synthesized and coated with collagen and stem cells were seeded on them for 24 hr. The obtained data showed that cells can attach faster to hydrogels compared with culture plate and cells on <1 kPa stiffness show more neuronal‐like morphology as they presented several branches and extended longer neurites over time. Moreover, the transcription of actin downregulated on all hydrogels, while the expression of Nestin, Tubulin, and PDGFR‐α increased on all of them and SOX‐10 and doublecortin gene expression were higher only on <1 kPa. Also, it was revealed that soft hydrogels can enhance the expression of glial cell line‐derived neurotrophic factor, neurotrophin‐3, and vascular endothelial growth factor in these stem cells. On the basis of the results, these cells can respond to the substrate stiffness in the short term culture and soft hydrogels can alter their morphology and gene expression. These findings suggested that employing proper substrate stiffness might result in cells with more natural profiles similar to the nervous system and superior usefulness in therapeutic applications.

Multi-Functionalized Self-Assembling Peptides as Reproducible 3D Cell Culture Systems Enabling Differentiation and Survival of Various Human Neural Stem Cell Lines

“…Moreover, Nestin positive cells decreased over time for all cell lines except for hNSC#1: this is likely ascribable to an initially higher proliferation rate (until 1WIV) caused by an enriched percentage of stem/progenitor cells population that progressively faded into differentiating (or fate-committed) cells ( Figure 4G). To note, the obtained values can be easily modified with further customizations of HYDROSAP depending on the regenerative application: indeed we already demonstrated that by tuning different key experimental parameters (e.g., matrix stiffness, presence of functional motifs, and/or addition of neurotrophic factors) it is possible to modulate NSCs differentiation and improve differentiation toward neuronal fate (Jain et al, 2020): for example, we showed that increments of cell proliferation rate and neuronal differentiation is inversely proportional to the matrix stiffness, while stiffness and astrocytes number directly correlated (Cunha et al, 2011;Caprini et al, 2013;Han et al, 2020;Pandamooz et al, 2020). In future experiments, it will also be interesting to assess the influence of time-evolving viscoelastic properties of hydrogels over cellular behavior (Mattei et al, 2020).…”

The Beneficial Potential of Genetically Modified Stem Cells in the Treatment of Stroke: a Review

“…This advantage over BM-MSCs supports the application of these stem cells to elderly individuals, who have an increased incidence of stroke. According to accumulating evidence, EPI-NCSCs express several trophic factors, which their rate of expression can be manipulated through the use of different preconditioning strategies [125][126][127][128][129] to achieve optimum efficiency after transplantation. In addition, administration of EPI-NCSCs via intra-arterial or intra venous routes following reperfusion, created a comparable outcome to intra-arterial grafted BM-MSCs, 7 days after cerebral ischemia [37].…”

The Beneficial Potential of Genetically Modified Stem Cells in the Treatment of Stroke: A Review

“…This advantage over BM-MSCs supports the application of these stem cells to elderly individuals, who have an increased incidence of stroke. According to accumulating evidence, EPI-NCSCs express several trophic factors, which their rate of expression can be manipulated through the use of different chemical agents [119][120][121][122] to achieve optimum efficiency after transplantation. In addition, administration of EPI-NCSCs via intra-arterial or intra venous routes following reperfusion, created a comparable outcome to intra-arterial grafted BM-MSCs, 7 days after cerebral ischemia [34].…”