Rahimi-Sherbaf Fatemeh, Nadri Samad, Rahmani Ali, Dabiri Oskoei Atousa
Department of Obstetrics and Gynecology, School of Medicine, Yas Hospital, Tehran University of Medical Sciences, Tehran, Iran.
Department of Medical Nanotechnology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.
Bioimpacts. 2020;10(2):117-122. doi: 10.34172/bi.2020.14. Epub 2020 Mar 26.
Transplantation of stem cells with a nanofibrous scaffold is a promising approach for spinal cord injury therapy. The aim of this work was to differentiate neural-like cells from placenta-derived mesenchymal stem cells (PDMSCs) using suitable induction reagents in three (3D) and two dimensional (2D) culture systems. After isolation and characterization of PDMSCs, the cells were cultivated on poly-L-lactide acid (PLLA)/poly caprolactone (PCL) nanofibrous scaffold and treated with a neuronal medium for 7 days. Electron microscopy, qPCR, and immunostaining were used to examine the differentiation of PDMSCs (on scaffold and tissue culture polystyrene [TCPS]) and the expression rate of neuronal markers (beta-tubulin, nestin, GFAP, and MAP-2). qPCR analysis showed that beta-tubulin (1.672 fold; ≤ 0.0001), nestin (11.145 fold; ≤ 0.0001), and GFAP (80.171; ≤ 0.0001) gene expressions were higher on scaffolds compared with TCPS. Immunofluorescence analysis showed that nestin and beta-tubulin proteins were recognized in the PDMSCs differentiated on TCPS and scaffold after 7 days in the neuroinductive differentiation medium. Taken together, these results delegated that PDMSCs differentiated on PLLA/PCL scaffolds are more likely to differentiate towards diversity lineages of neural cells. It proposed that PDMSCs have cell subpopulations that have the capability to be differentiated into neurogenic cells.
将干细胞与纳米纤维支架一起移植是脊髓损伤治疗的一种有前景的方法。这项工作的目的是在三维（3D）和二维（2D）培养系统中使用合适的诱导试剂，使胎盘来源的间充质干细胞（PDMSCs）分化为神经样细胞。在对PDMSCs进行分离和鉴定后，将细胞接种在聚-L-乳酸（PLLA）/聚己内酯（PCL）纳米纤维支架上，并用神经元培养基处理7天。使用电子显微镜、qPCR和免疫染色来检测PDMSCs（在支架和组织培养聚苯乙烯[TCPS]上）的分化情况以及神经元标志物（β-微管蛋白、巢蛋白、胶质纤维酸性蛋白和微管相关蛋白2）的表达率。qPCR分析表明，与TCPS相比，支架上β-微管蛋白（1.672倍；≤0.0001）、巢蛋白（11.145倍；≤0.0001）和胶质纤维酸性蛋白（80.171；≤0.0001）的基因表达更高。免疫荧光分析表明，在神经诱导分化培养基中培养7天后，在TCPS和支架上分化的PDMSCs中可检测到巢蛋白和β-微管蛋白。综上所述，这些结果表明在PLLA/PCL支架上分化的PDMSCs更有可能向神经细胞的不同谱系分化。这表明PDMSCs具有能够分化为神经源性细胞的细胞亚群。
