Differentiation of Wharton’ s Jelly-Derived Mesenchymal Stem Cells into Motor Neuron-Like Cells on Three-Dimensional Collagen-Grafted Nanofibers Zohreh Bagher 1 & Mahmoud Azami 2 & Somayeh Ebrahimi-Barough 2,3 & Hamid Mirzadeh 4 & Atefeh solouk 5 & Mansooreh Soleimani 1,6,7 & Jafar Ai 2,3 & Mohammad Reza Nourani 8 & Mohammad Taghi Joghataei 1,6,7 Received: 14 December 2014 /Accepted: 27 April 2015 # Springer Science+Business Media New York 2015 Abstract Cell transplantation strategies have provided poten- tial 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 trans- plantation 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 proper- ties of scaffolds have been shown to significantly influence cell behaviors such as adhesion, proliferation, and differentiation. Therefore, polycaprolactone (PCL) nanofibers were construct- ed 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 morphologi- cally, 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. Keywords Wharton’ s Jelly-derived mesenchymal stem cell . Motor neuron differentiation . Nanofibrous scaffolds Introduction Spinal cord injury (SCI) often leads to cell death and axonal degeneration resulting in functional motor and sensory loss [1]. In many cases, spinal cord is unable to regenerate or repair its functions following pathological trauma or disease [2–4]. * Mohammad Taghi Joghataei joghataei.mt@iums.ac.ir 1 Department of Tissue Engineering and Regenerative Medicine, School of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran 2 Department of Tissue Engineering and Regenerative Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran 3 Brain and Spinal Injury Research Center, Imam Khomeini Hospital, Tehran University of Medical Sciences, Tehran, Iran 4 Polymer Engineering Faculty, Amirkabir University of Technology (Tehran Polytechnic), Tehran, Iran 5 Biomedical Engineering Faculty, Amirkabir University of Technology (Tehran Polytechnic), Tehran, Iran 6 Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran 7 Department of Anatomy, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran 8 Tissue Engineering Division, Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran Mol Neurobiol DOI 10.1007/s12035-015-9199-x