Differentiation of Wharton's Jelly-Derived Mesenchymal Stem Cells into Motor Neuron-Like Cells on Three-Dimensional Collagen-Grafted Nanofibers
Differentiation of Wharton's Jelly-Derived Mesenchymal Stem Cells into Motor Neuron-Like Cells on Three-Dimensional Collagen-Grafted Nanofibers
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DOI：
10.1007/s12035-015-9199-x
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发表时间：
2016-05-01
影响因子：
5.1
通讯作者：
Joghataei, Mohammad Taghi
中科院分区：
文献类型：
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作者：
Bagher, Zohreh;Azami, Mahmoud;Joghataei, Mohammad Taghi
细胞移植策略为神经退行性疾病的治疗提供了潜在的治疗方法。华顿胶间充质干细胞（Mesenchymal stem cells from Wharton's jelly，WJMSCs）是一种来源丰富、免疫相容性较低的成体干细胞，可作为细胞替代治疗的候选细胞。然而，没有任何诱导或支持材料的MSC移植导致细胞活力和分化的控制不良。在这项研究中，我们研究了在维甲酸（RA）和音刺猬（Shh）存在下，纳米支架对WJMSCs分化为运动神经元谱系的影响。支架的表面性质已经显示出显著影响细胞行为，例如粘附、增殖和分化。因此，聚己内酯（PCL）纳米纤维的构建通过静电纺丝，表面改性的等离子体处理，并接枝胶原蛋白。通过ATR-FTIR、接触角和Bradford表征支架，证明胶原在支架表面接枝。将WJMSCs接种在纳米纤维和组织培养板（TCP）上，通过MTT法测定WJMSCs的活力，然后诱导WJMSCs分化为运动神经元样细胞15天。对分化后的细胞进行形态学观察，并采用实时荧光定量PCR和免疫细胞化学方法检测运动神经元样细胞标志物在mRNA和蛋白水平的表达。我们的结果表明，获得的细胞可以表达运动神经元生物标志物在RNA和蛋白质水平，但WJMSCs的生存和分化为运动神经元样细胞的PCL/胶原支架上比TCP和PCL组培养的细胞。总之，WJMSC是一种有吸引力的干细胞来源，用于在体外诱导成运动神经元，特别是当在纳米结构支架上生长时，并且PCL/胶原支架可以为神经元存活和分化提供合适的三维情况，这表明它们对神经再生的潜在应用。
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.