Wang Tony Y, Sen Arindom, Behie Leo A, Kallos Michael S
Pharmaceutical Production Research Facility (PPRF), Schulich School of Engineering, University of Calgary, 2500 University Drive N.W., Calgary, Alberta, Canada T2N 1N4.
Brain Res. 2006 Aug 30;1107(1):82-96. doi: 10.1016/j.brainres.2006.05.110. Epub 2006 Jul 21.
Large-scale expansion of neural stem and progenitor cells will be essential for clinically treating the large number of patients suffering from neurodegenerative disorders such as Parkinson's disease. Other applications of neural stem cell technology include further research in areas such as neural development or drug testing. Neural stem cells can be grown in vitro as tissue aggregates known as neurospheres, and in the current study, experiments were performed to determine the spatial arrangement and behavior of the cells within the neurosphere structure. A protocol utilizing sulfonated lipophilic fluorescent dyes was developed to effectively label populations of neural stem and progenitor cells without compromising cell density during culture. Cells retained the labels for at least 7 days. Using the labeling protocol, we discovered that the cells within the neurospheres were mobile and, moreover, the cells on the periphery of the neurospheres could migrate into the center of the neurospheres. Most important, the mixing time of two merging neurospheres was observed to be the same order of magnitude as the neural stem cell doubling time (approximately 20 h). This study is the first to show that the neurosphere system is dynamic, and these results will serve as a stepping stone to more in-depth studies of the neurosphere microenvironment.
大规模扩增神经干细胞和祖细胞对于临床上治疗大量患有神经退行性疾病（如帕金森病）的患者至关重要。神经干细胞技术的其他应用包括在神经发育或药物测试等领域的进一步研究。神经干细胞可以在体外作为称为神经球的组织聚集体生长，在当前的研究中，进行了实验以确定神经球结构内细胞的空间排列和行为。开发了一种利用磺化亲脂性荧光染料的方案，以有效地标记神经干细胞和祖细胞群体，同时在培养过程中不影响细胞密度。细胞至少保留标记7天。使用该标记方案，我们发现神经球内的细胞是可移动的，此外，神经球外围的细胞可以迁移到神经球的中心。最重要的是，观察到两个合并神经球的混合时间与神经干细胞倍增时间（约20小时）处于相同的数量级。这项研究首次表明神经球系统是动态的，这些结果将作为对神经球微环境进行更深入研究的垫脚石。
