Coseeded Schwann cells myelinate neurites from differentiated neural stem cells in neurotrophin-3-loaded PLGA carriers
Coseeded Schwann cells myelinate neurites from differentiated neural stem cells in neurotrophin-3-loaded PLGA carriers
复制标题
DOI：
10.2147/ijn.s30706
复制
发表时间：
2012-04
影响因子：
8
通讯作者：
Y. Xiong;Ji-xiang Zhu;Z. Fang;Cheng Zeng;Chao Zhang;Guo-long Qi;Manhui Li;Wei Zhang;D. Quan
中科院分区：
文献类型：
--
作者：
Y. Xiong;Ji-xiang Zhu;Z. Fang;Cheng Zeng;Chao Zhang;Guo-long Qi;Manhui Li;Wei Zhang;D. Quan
生物材料和神经营养因子代表了神经修复的有希望的指导。在这项研究中，我们结合聚（乳酸-羟基乙酸）（PLGA）导管和神经营养素-3（NT-3），以产生NT-3负载PLGA载体在体外。生物活性NT-3从PLGA导管中稳定和持续释放长达4周。将神经干细胞（NSC）和雪旺细胞（SC）共接种到NT释放支架系统中并培养14天。对Map 2的免疫反应性显示，大多数移植细胞（>80%）向神经元分化。突触发生和髓鞘形成的双重免疫组化染色显示，在共培养中形成了突触结构和髓鞘，这也在电子显微镜下观察到。此外，在去极化条件下，这些突触是可兴奋的，并能够释放与FM 1 -43或FM 4 -64标记的突触囊泡。两者合计，共接种神经干细胞和SC到NT-3负载的PLGA载体增加了神经干细胞分化成神经元，开发突触连接，表现出突触活动，和伴随的SC的神经突髓鞘形成。这些结果为脊髓损伤后功能性神经重建的移植提供了实验依据。
Biomaterials and neurotrophic factors represent promising guidance for neural repair. In this study, we combined poly-(lactic acid-co-glycolic acid) (PLGA) conduits and neurotrophin-3 (NT-3) to generate NT-3-loaded PLGA carriers in vitro. Bioactive NT-3 was released stably and constantly from PLGA conduits for up to 4 weeks. Neural stem cells (NSCs) and Schwann cells (SCs) were coseeded into an NT-releasing scaffold system and cultured for 14 days. Immunoreactivity against Map2 showed that most of the grafted cells (>80%) were differentiated toward neurons. Double-immunostaining for synaptogenesis and myelination revealed the formation of synaptic structures and myelin sheaths in the coculture, which was also observed under electron microscope. Furthermore, under depolarizing conditions, these synapses were excitable and capable of releasing synaptic vesicles labeled with FM1-43 or FM4-64. Taken together, coseeding NSCs and SCs into NT-3-loaded PLGA carriers increased the differentiation of NSCs into neurons, developed synaptic connections, exhibited synaptic activities, and myelination of neurites by the accompanying SCs. These results provide an experimental basis that supports transplantation of functional neural construction in spinal cord injury.