摘要
目的利用硫酸软骨素蛋白多糖(CSPG)的生物特性建立抑制神经元生长和轴突再生的体外生物模型。方法将不同浓度CSPG与无生物毒性的德克萨斯红荧光染料混合,取5μl滴在培养板的中心部位,形成边界清楚的红色圆形斑点,称CSPG圆斑(CSPG SPOT)。将神经母细胞瘤SY5Y细胞和小鼠小脑颗粒细胞(CGNs)铺在含有CSPG SPOT的培养板中,48h后观察细胞的生长情况。结果含有1μg/ml和3μg/ml CSPG的SPOT分别使CGNs和神经母细胞瘤SY5Y细胞停止生长、死亡,没有发现细胞的轴突延伸到SPOT上生长。高浓度的CSPG可渗透到SPOT周围区域,使CSPG SPOT周围区域的细胞生长和轴突再生受到抑制。结论该模型可以在体外模拟胶质细胞分泌的CSPG对神经细胞生长的抑制作用,对研究中枢神经系统损伤后神经元的生长和轴突的再生具有广泛的应用前景。
Objective To establish an in vitro model to study the inhibitory effect of chondroitin sulfate proteoglycan （CSPG） on neuronal growth and axonal regeneration.Methods Different concentrations of CSPG were mixed with Texas-red Dye which has no biological toxicity,and 5 μl of the mixture was dropped onto the center of the bottom of culture plates to form red round spots,which was called CSPG spots.SY5Y neuroblastoma cells or cerebellar granule neuron （CGN） from mice were seeded on the plates and cultured for 48 hours.The cell growth and axon extension were observed under microscope.Results In CSPG spots containing 1 and 3 μg/ml of CSPG,the growth of CGNs and SY5Y neuroblastoma cells were inhibited,and cell death occurred in the spot areas.No axon extension was found in cells outside the CSPG spots.High concentration of CSPG in the spots inhibited the growth of cells around CSPG spots.Conclusion This model mimics the CSGP-induced axon outgrowth inhibition,which can be used to study the neuronal growth and axonal regeneration after central nervous system injury.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2011年第1期4-7,11,共5页
Journal of China Medical University
基金
辽宁省科技厅社发基金资助项目(2009225010-12)
