Pandamooz Sareh, Naji Mohammad, Alinezhad Farid, Zarghami Amin, Pourghasem Mohsen
Stem Cell Res Ther. 2013 Jul 18;4(4):84. doi: 10.1186/scrt235.
Epidermal neural crest stem cells (EPI-NCSCs) in the bulge of hair follicles are a promising source for cell-replacement therapies in neurodegenerative diseases. A prominent factor in cell-based therapy is the practicalities of different routes of administration. Cerebrospinal fluid (CSF), owing to its adaptive library of secreted growth factors, can provide a trophic environment for transplanted cells. Thus, the effect of CSF on the behavior of EPI-NCSC was studied here.
In this study, the highly pure population of EPI-NCSCs was obtained from the bulge of mouse hair follicle. Migrated cells were characterized with real-time polymerase chain reaction (RT-PCR) and immunocytochemistry. Subsequently isolated stem cells were cultured in CSF, which was collected from the cisterna magna of the adult rat. The expression of pertinent markers was assessed at the gene and protein levels with RT-PCR and immunocytochemistry, respectively. Colorimetric immunoassay was used to quantify the rate of proliferation of EPI-NCSCs after cultivation in CSF.
Isolated EPI-NCSCs could survive in the CSF, and they maintained the expression of nestin, β-tubulin ІІІ (early neuronal marker), and glial fibrillary acidic protein (GFAP, glia marker) in this environment. In addition, CSF decreased the proliferation rate of EPI-NCSCs significantly in comparison to primary and expansion culture medium.
Our findings demonstrate that CSF as a cocktail of growth factors helps EPI-NCSCs to acquire some desirable traits, and because of its circulatory system that is in close contact with different parts of the central nervous system (CNS), can be a practical route of administration for delivery of injected stem cells.
毛囊隆突部的表皮神经嵴干细胞(EPI-NCSCs)是神经退行性疾病细胞替代疗法的一个有前景的细胞来源。基于细胞的治疗中的一个重要因素是不同给药途径的实际应用。脑脊液(CSF)因其分泌的生长因子的适应性库,可为移植细胞提供营养环境。因此,本研究探讨了脑脊液对EPI-NCSC细胞行为的影响。
在本研究中,从小鼠毛囊隆突部获得了高度纯化的EPI-NCSCs群体。通过实时聚合酶链反应(RT-PCR)和免疫细胞化学对迁移的细胞进行表征。随后将分离的干细胞培养在从成年大鼠小脑延髓池收集的脑脊液中。分别用RT-PCR和免疫细胞化学在基因和蛋白质水平评估相关标志物的表达。采用比色免疫分析法量化EPI-NCSCs在脑脊液中培养后的增殖率。
分离的EPI-NCSCs能够在脑脊液中存活,并且在这种环境中维持巢蛋白、β-微管蛋白III(早期神经元标志物)和胶质纤维酸性蛋白(GFAP,神经胶质标志物)的表达。此外,与原代和传代培养基相比,脑脊液显著降低了EPI-NCSCs的增殖率。
我们的研究结果表明,脑脊液作为一种生长因子混合物有助于EPI-NCSCs获得一些理想的特性,并且由于其与中枢神经系统(CNS)不同部位紧密接触的循环系统,可成为注射干细胞递送的一种实用给药途径。