Differentiation of Wharton’s Jelly-Derived Mesenchymal Stem Cells into Motor Neuron-Like Cells on Three-Dimensional Collagen-Grafted Nanofibers

Abstract

Cell transplantation strategies have provided potential therapeutic approaches for treatment of neurodegenerative diseases. Mesenchymal stem cells from Wharton's jelly (WJMSCs) are abundant and available adult stem cells with low immunological incompatibility, which could be considered for cell replacement therapy in the future. However, MSC transplantation without any induction or support material causes poor control of cell viability and differentiation. In this study, we investigated the effect of the nanoscaffolds on WJMSCs differentiation into motor neuronal lineages in the presence of retinoic acid (RA) and sonic hedgehog (Shh). Surface properties of scaffolds have been shown to significantly influence cell behaviors such as adhesion, proliferation, and differentiation. Therefore, polycaprolactone (PCL) nanofibers were constructed via electrospinning, surface modified by plasma treatment, and grafted by collagen. Characterization of the scaffolds by means of ATR-FTIR, contact angel, and Bradford proved grafting of the collagen on the surface of the scaffolds. WJMSCs were seeded on nanofibrous and tissue culture plate (TCP) and viability of WJMSCs were measured by MTT assay and then induced to differentiate into motor neuron-like cells for 15 days. Differentiated cells were evaluated morphologically, and real-time PCR and immunocytochemistry methods were done to evaluate expression of motor neuron-like cell markers in mRNA and protein levels. Our results showed that obtained cells could express motor neuron biomarkers at both RNA and protein levels, but the survival and differentiation of WJMSCs into motor neuron-like cells on the PCL/collagen scaffold were higher than cultured cells in the TCP and PCL groups. Taken together, WJMSCs are an attractive stem cell source for inducing into motor neurons in vitro especially when grown on nanostructural scaffolds and PCL/collagen scaffolds can provide a suitable, three-dimensional situation for neuronal survival and differentiation that suggest their potential application towards nerve regeneration.

Cellular activity of Wharton's Jelly‐derived mesenchymal stem cells on electrospun fibrous and solvent‐cast film scaffolds

“…WJMSCs were induced to motor neuron‐like cells according to a previously reported protocol . For motor neuron‐like cell induction, 2 × 10 5 cells/cm 2 were seeded on the scaffolds and let them to grow for 24 h in previously defined media.…”

Nanotechnology for mesenchymal stem cell therapies

“…Accordingly, random nanofibers have been recently reported to significantly improve rat MSC proliferation compared to aligned nanofibers [99]. The [96] or in combination with sonic Hedgehog [105], graphene oxide [106], bioceramics [107], growth factors [108], acellular pericardium [109], platelet-rich plasma [110], collagen [104], fibronectin [111], and mussel inspired DOPA-melanin coating for the adsorption of siRNA targeting RE-1 silencing transcription factor [112]. In all cases, depending on the desired application, nanostructural surfaces (whether scaffolds or membranes) have been reported to provide a suitable system to support adhesion and growth of the cell type of interest, and to influence progenitor population activity and function, holding promise for tissue regeneration applications.…”

Current status of stem cell therapy: opportunities and limitations

“…The use of stem cells in neural differentiation/ neuroregeneration was also reported, such as for induction of motor neuron differentiation applied with complex matrices that included collagen grafted nanofibers (Bagher et al, 2015), or peripheral nerve repair with single-walled carbon nanotubes/poly-lactic acid scaffolds (Kabiri et al, 2015).…”