Liu I-Hsien, Chen Shih-Jen, Ku Hung-Hai, Kao Chung-Lan, Tsai Fu-Ting, Hsu Wen-Ming, Lo Chih-Wen, Kuo Yao-Haur, Kuo Cheng-Deng, Lee Chen-Hsen, Chiou Shih-Hwa
Department of Ophthalmology, Taipei Veterans General Hospital and National Yang-Ming University, Taipei, Taiwan, ROC.
Ophthalmologica. 2005 May-Jun;219(3):171-6. doi: 10.1159/000085250.
Recent studies have demonstrated that retinal stem cells (RSCs) and stem cells of the central nervous system both exhibited the abilities of self-renewal, proliferation and differentiation into multilineage. In the present study, we compared the proliferation and differentiation abilities between RSCs and cerebral corticex-derived neural stem cells (CNSCs) of adult rats. Stem cells isolated from pigmented ciliary margins of eyes and cerebral cortical tissues of adult rats were cultured in 96-well plates that contained serum-free medium with epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF). In contrast to RSCs, which stopped proliferating after the 8th week, the total cell count of neurospheres in CNSCs increased twofold at the 5th week and more than fourfold at the 10th week after in vitro culture. In contrast, RSCs stopped proliferating after 8 weeks of culture. After adding 2% fetal calf serum and withdrawing EGF and bFGF from the culture medium, the percentages of nestin-positive cells(20.6 +/- 2.7%), microtubule-associated-protein-2-positive neurons (33.2 +/- 3.9%) and glial-fibrillary-acidic-protein-positive glial cells(51.3 +/- 6.2%) in the differentiated CNSCs were significantly higher than those in the differentiated RSCs (10.2 +/- 1.9, 22.3 +/- 1.3 and 44.6 +/- 5.1%, respectively; p < 0.05). We also found that the combination of transforming growth factor beta type III with retinoic acid played an important role in the induction of CNSCs to differentiate into opsin-positive cells. Our data demonstrated that CNSCs displayed a higher ability of proliferation and retinal lineage. This report also offers an alternative protocol of cell reproduction for producing retinal cells.
近期研究表明，视网膜干细胞（RSCs）和中枢神经系统干细胞均具有自我更新、增殖以及分化为多谱系细胞的能力。在本研究中，我们比较了成年大鼠RSCs与大脑皮质来源的神经干细胞（CNSCs）的增殖和分化能力。从成年大鼠眼色素睫状缘和大脑皮质组织分离出的干细胞，在含有表皮生长因子（EGF）和碱性成纤维细胞生长因子（bFGF）的无血清培养基的96孔板中培养。与RSCs不同，RSCs在第8周后停止增殖，而CNSCs的神经球总细胞数在体外培养第5周时增加了两倍，在第10周时增加了四倍多。相比之下，RSCs在培养8周后停止增殖。在向培养基中添加2%胎牛血清并去除EGF和bFGF后，分化的CNSCs中巢蛋白阳性细胞（20.6±2.7%）、微管相关蛋白2阳性神经元（33.2±3.9%）和胶质纤维酸性蛋白阳性胶质细胞（51.3±6.2%）的百分比显著高于分化的RSCs（分别为10.2±1.9、22.3±1.3和44.6±5.1%；p<0.05）。我们还发现，转化生长因子βⅢ型与视黄酸的组合在诱导CNSCs分化为视蛋白阳性细胞中起重要作用。我们的数据表明，CNSCs表现出更高的增殖能力和视网膜谱系分化能力。本报告还提供了一种用于产生视网膜细胞的细胞增殖替代方案。
