Functional surfaces for efficient differentiation of neural stem/progenitor cells into dopaminergic neurons

Abstract

Various techniques and systems have been reported for the efficient differentiation of neural stem/progenitor cells into dopaminergic neurons. Although a comparatively high percentage of dopaminergic neurons can be obtained using these techniques, the differentiated cells display varied cellular phenotypes such as astrocytes and oligodendrocytes. Generation of highly pure dopaminergic neurons is important for cell‐based therapy and in vitro evaluation of dopaminergic neuron function. In this study, we developed a culture surface anchored with several neurotrophic factors and a neuronal cell‐adhesive protein for efficient differentiation of neural stem/progenitor cells into dopaminergic neurons. Oligohistidine‐fused brain‐derived neurotrophic factor and glial cell line–derived neurotrophic factor, synthesized using genetic engineering, were co‐immobilized on the surface via metal chelation. To facilitate cell adhesion, a cell‐adhesive chimeric protein derived from laminin (LN‐G) was also immobilized on the surface. Approximately 40% of the cells cultured for 14 days with these protein‐immobilized substrates expressed tyrosine hydroxylase, a marker of dopaminergic neurons, with a three‐fold increase in differentiation efficiency than that reported previously. In addition, the number of tyrosine hydroxylase‐positive cells increased to approximately 80% of the culture after 30 days. These cells secreted dopamine and expressed dopaminergic neuron‐specific genes. Interestingly, cell types (glial cells and oligodendrocytes) other than neuronal cells (immature and mature dopaminergic neurons) were not detected on the protein‐anchored surface. Our results demonstrate that highly pure dopaminergic neurons can be exclusively obtained using the novel substrate without extra purification steps such as cell sorting. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 860–871, 2019.

Divergence between Neuronal and Oligodendroglial Cell Fate, in Postnatal Brain Neural Stem Cells, Leads to Divergent Properties in Polymorphic In Vitro Assays

“…Experimental work on extracellular matrix scaffolds and NSC differentiation has produced contradicting results on NSC-derived oligodendrogenesis, indicating the involvement of intricate and different mechanisms. One study reported that when NSCs were cultured in a methylcellulose scaffold functionalized with laminin, OPC generation was increased [ 48 ], while co-immobilizing laminin with several neurotrophic factors on a culture surface inhibited this process [ 49 ].…”

3D printed scaffolds based on hyaluronic acid bioinks for tissue engineering: a review

“…These methods have been used to design HA-based polymers with ideal rheological properties for 3D printing, and they do not affect the nucleophilic nature of the permanent exchange of sulfhydryl groups with disulfides. A supramolecular hydrogel based on HA and recombinant peptides was prepared using metal–ligand bonding to enhance the capture and release performance of bioactive constituents [ 114 ].…”

Neural stem cells-from quiescence to differentiation and potential clinical uses

“…This treatment led to elevated expression of dopaminergic factors like Otx2 and Msh homeobox 1 (Msx1) along with tyrosine hydroxylase and pituitary homeobox 3 (Pitx3) (Rössler et al, 2010). Other studies also utilised B27, GDNF and BDNF to induce dopaminergic differentiation (Alizadeh et al, 2017;Nakaji-Hirabayashi et al, 2019;Daadi 2019). An important role is attributed to p70 ribosomal S6 kinase (S6K), a downstream effector of PI3K/Akt/mTOR signaling that is activated in response to insulin in dopaminergic neurons (Fang et al, 2007).…”