Xiong Yi, Zeng Yuan-Shan, Zeng Chen-Guang, Du Bao-Ling, He Liu-Min, Quan Da-Ping, Zhang Wei, Wang Jun-Mei, Wu Jin-Lang, Li Yan, Li Jun
Division of Neuroscience, Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China.
Biomaterials. 2009 Aug;30(22):3711-22. doi: 10.1016/j.biomaterials.2009.03.046. Epub 2009 Apr 17.
To explore therapeutic potential of engineered neural tissue, we combined genetically modified neural stem cells (NSCs) and poly(lactic acid-co-glycolic acid) (PLGA) polymers to generate an artificial neural network in vitro. NSCs transfected with either NT-3 or its receptor TrkC gene were seeded into PLGA scaffold. The NSCs were widely distributed and viable in the scaffold after culturing for 14 days. Immunoreactivity against Map2 was detected in >70% of these grafted cells, suggesting a high rate of differentiation toward neurons. Immunostaining of synapsin-I and PSD95 revealed formation of synaptic structures, which was also observed under electron microscope. Furthermore, using FM1-43 dynamic imaging, synapses in these differentiated neurons were found to be excitable and capable of releasing synaptic vesicles. Taken together, our artificial PLGA construct permits NSCs to differentiate toward neurons, establish connections and exhibit synaptic activities. These findings provide a biological basis for future application or transplantation of this artificial construct in neural repair.
为了探索工程化神经组织的治疗潜力，我们将基因改造的神经干细胞（NSCs）与聚乳酸-乙醇酸共聚物（PLGA）相结合，在体外构建了一个人工神经网络。将转染了NT-3或其受体TrkC基因的神经干细胞接种到PLGA支架中。培养14天后，神经干细胞在支架中广泛分布且存活。在超过70%的这些移植细胞中检测到了针对Map2的免疫反应性，表明向神经元分化的比例很高。突触素-I和PSD95的免疫染色显示形成了突触结构，在电子显微镜下也观察到了这一现象。此外，使用FM1-43动态成像发现，这些分化神经元中的突触具有兴奋性，能够释放突触小泡。综上所述，我们的人工PLGA构建体使神经干细胞能够向神经元分化、建立连接并表现出突触活动。这些发现为这种人工构建体未来在神经修复中的应用或移植提供了生物学基础。
