Ebrahimi-Barough Somayeh, Hoveizi Elham, Norouzi Javidan Abbas, Ai Jafar
Brain and Spinal Cord Injury Research Center (BASIR), Tehran University of Medical Sciences, Keshavarz Boulevard, Gharib Street, Tehran, 6114185, Iran.
Department of Tissue Engineering and Applied Cell Sciences, Faculty of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.
J Biomed Mater Res A. 2015 Aug;103(8):2621-7. doi: 10.1002/jbm.a.35397. Epub 2015 Feb 2.
Neural tissue engineering is an important area of research in the field of tissue-engineering especially for neurodegenerative disease such as spinal cord injury. The differentiation capacity of human endometrial stem cells (hEnSCs) into neuronal cells has yet to be elucidated. Here, the major aim of the present study was to investigate the differentiation ability of hEnSCs cultured on polylactic acid/chitosan (PLA/CS) nanofibrous scaffold into neuroglial cells in response to conditioned medium of BE(2)-C human neuroblastoma cells and growth factors. Here we investigated the use PLA/CS scaffold as a three dimensional (3D) system that increased neuro-glial cells differentiation. Human EnSCs after three passages were differentiated in neuro-glial like cells under neuroblastoma conditioned medium with FGF2/PDGF-AA on PLA/CS scaffold. By day 18, differentiated cells were analyzed for expression of neuroglial markers by qRT-PCR and immunofluorescence. The results revealed that hEnSCs attach, grow and differentiation on the nanofibrous PLA/CS scaffold. Additionally, our study showed the expression of neural and glial lineage markers such as Nestin, NF-L, MAP2, PDGFRa, CNP, Olig2, MBP, and GFAP in the level of mRNA and MAP2, Tuj-1, and NF-L in the protein level after 18 days. Our results demonstrate that hEnSCs cultured on PLA/CS nanofibrous scaffold have the potential to differentiate in neuronal and glial cells in presence of neuroblastoma conditioned medium on PLA/CS scaffold. The result of this study may have impact in tissue engineering and cells-base therapy of neurodegenerative diseases and have a great potential for wide application.
神经组织工程是组织工程领域的一个重要研究方向，尤其对于诸如脊髓损伤等神经退行性疾病而言。人子宫内膜干细胞（hEnSCs）向神经元细胞的分化能力尚未阐明。在此，本研究的主要目的是探究在聚乳酸/壳聚糖（PLA/CS）纳米纤维支架上培养的hEnSCs，在人神经母细胞瘤BE(2)-C细胞条件培养基和生长因子作用下向神经胶质细胞的分化能力。在此，我们研究了使用PLA/CS支架作为一种三维（3D）系统，该系统可促进神经胶质细胞的分化。传代三次后的人EnSCs在PLA/CS支架上，于神经母细胞瘤条件培养基中添加FGF2/PDGF-AA的情况下分化为神经胶质样细胞。在第18天时，通过qRT-PCR和免疫荧光分析分化细胞中神经胶质标志物的表达。结果显示，hEnSCs可附着、生长并在纳米纤维PLA/CS支架上分化。此外，我们的研究表明，18天后，在mRNA水平上有神经和胶质谱系标志物如巢蛋白、神经丝轻链（NF-L）、微管相关蛋白2（MAP2）、血小板衍生生长因子受体α（PDGFRα）、2',3'-环核苷酸3'-磷酸二酯酶（CNP）、少突胶质细胞转录因子2（Olig2）、髓鞘碱性蛋白（MBP）和胶质纤维酸性蛋白（GFAP）表达，在蛋白水平上有MAP2、βIII微管蛋白（Tuj-1）和NF-L表达。我们的结果表明，在PLA/CS支架上，于神经母细胞瘤条件培养基存在的情况下，在PLA/CS纳米纤维支架上培养的hEnSCs有向神经元和胶质细胞分化的潜力。本研究结果可能对神经退行性疾病的组织工程和细胞基治疗产生影响，具有广泛应用的巨大潜力。
