Ghoroghi Fatemeh Moghani, Hejazian Leila Beygom, Esmaielzade Banafshe, Dodel Masumeh, Roudbari Masoud, Nobakht Maliheh
Anatomy Department, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
J Mol Neurosci. 2013 Aug 20. doi: 10.1007/s12031-013-0073-x.
Recent improvement in neuroscience has led to new strategies in neural repair. Hair follicle stem cells are high promising source of accessible, active, and pluripotent adult stem cells. They have high affinity to differentiate to neurons. Aside from using cell-scaffold combinations for implantation, scaffolds can provide a suitable microenvironment for cell proliferation, migration, and differentiation. NT-3 is the most interesting neurotrophic factors being an important regulator of neural survival and differentiation. Since treatment duration in neural repair is very important, this study aims to evaluate the effect of NT-3 and poly-L-lactic acid (PLLA) on differentiation time of bulge stem cells of rat hair follicle to neural-like cells. HFSCs of rat whisker was isolated and cultured on PLLA and differentiated with 10 ng/mL NT-3. Biological features of cultured cells were evaluated with immunocytochemistry and flowcytometry methods by using CD34, nestin, and βІІІ-tubulin markers. For cell viability and morphological assessment, MTT assay and SEM were performed. Our results showed that bulge stem cells of hair follicle can express CD34 and Nestin before differentiation. By using NT-3 during differentiation process, the cells showed positive reaction to βІІІ-tubulin antibody. MTT results demonstrated that PLLA significantly increased cell viability. Finally, HFSCs adhesion was confirmed by SEM results. The results indicate that 10 ng/mL NT-3 and PLLA have significant effect on differentiation time of rat HFSCs to neural cells even in 10 days.
神经科学领域的最新进展催生了神经修复的新策略。毛囊干细胞是可获取、活跃且具有多能性的成体干细胞的极有前景的来源。它们具有向神经元分化的高亲和力。除了使用细胞 - 支架组合进行植入外,支架可为细胞增殖、迁移和分化提供合适的微环境。NT - 3是最受关注的神经营养因子,是神经存活和分化的重要调节因子。由于神经修复中的治疗持续时间非常重要,本研究旨在评估NT - 3和聚 - L - 乳酸(PLLA)对大鼠毛囊隆突干细胞向神经样细胞分化时间的影响。分离大鼠触须的毛囊干细胞并在PLLA上培养,并用10 ng/mL NT - 3进行分化。通过使用CD34、巢蛋白和βⅢ - 微管蛋白标记物,采用免疫细胞化学和流式细胞术方法评估培养细胞的生物学特性。为了进行细胞活力和形态学评估,进行了MTT试验和扫描电子显微镜(SEM)检测。我们的结果表明,毛囊隆突干细胞在分化前可表达CD34和巢蛋白。在分化过程中使用NT - 3时,细胞对βⅢ - 微管蛋白抗体呈阳性反应。MTT结果表明PLLA显著提高了细胞活力。最后,SEM结果证实了毛囊干细胞的黏附。结果表明,即使在10天内,10 ng/mL NT - 3和PLLA对大鼠毛囊干细胞向神经细胞的分化时间也有显著影响。