Soliman Erfan, Bianchi Fabio, Sleigh James N, George Julian H, Cader M Zameel, Cui Zhanfeng, Ye Hua
Institute of Biomedical Engineering, Old Road Campus Research Building, University of Oxford, Oxford, OX3 7DQ, UK.
The Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital/Headley Way, Oxford, OX3 9DS, UK.
Biotechnol Lett. 2018 Mar;40(3):601-607. doi: 10.1007/s10529-017-2494-z. Epub 2018 Jan 8.
To test a 3D approach for neural network formation, alignment, and patterning that is reproducible and sufficiently stable to allow for easy manipulation.
A novel cell culture system was designed by engineering a method for the directional growth of neurons. This uses NG108-15 neuroblastoma x glioma hybrid cells cultured on suspended and aligned electrospun fibers. These fiber networks improved cellular directionality, with alignment angle standard deviations significantly lower on fibers than on regular culture surfaces. Morphological studies found nuclear aspect ratios and cell projection lengths to be unchanged, indicating that cells maintained neural morphology while growing on fibers and forming a 3D network. Furthermore, fibronectin-coated fibers enhanced neurite extensions for all investigated time points. Differentiated neurons exhibited significant increases in average neurite lengths 96 h post plating, and formed neurite extensions parallel to suspended fibers, as visualized through scanning electron microscopy.
The developed model has the potential to serve as the basis for advanced 3D studies, providing an original approach to neural network patterning and setting the groundwork for further investigations into functionality.
测试一种用于神经网络形成、排列和图案化的三维方法,该方法具有可重复性且足够稳定,便于操作。
通过设计一种神经元定向生长的方法,设计了一种新型细胞培养系统。该系统使用在悬浮且排列的电纺纤维上培养的NG108-15神经母细胞瘤x胶质瘤杂交细胞。这些纤维网络改善了细胞的方向性,纤维上的排列角度标准偏差明显低于常规培养表面。形态学研究发现细胞核长宽比和细胞突起长度没有变化,表明细胞在纤维上生长并形成三维网络时保持了神经形态。此外,在所有研究的时间点,纤连蛋白包被的纤维增强了神经突的延伸。分化的神经元在接种后96小时平均神经突长度显著增加,并形成了与悬浮纤维平行的神经突延伸,这通过扫描电子显微镜得以观察到。
所开发的模型有潜力作为高级三维研究的基础,为神经网络图案化提供一种原创方法,并为进一步的功能研究奠定基础。