Yi Xin, Jin Guohua, Tian Meiling, Mao Weifeng, Qin Jianbing
Department of Anatomy and Neurobiology, Nantong University, Nantong, China.
Neuro Endocrinol Lett. 2011;32(5):705-10.
Successful neural stem cells (NSCs) therapies require the controlled differentiation of NSCs into neurons. Porous chitosan scaffold was explored if it promoted neuronal differentiation of NSCs in the presence of nerve growth factor (NGF) in 3-dimensional (3-D) culture.
Chitosan scaffold was made by the freeze-drying technique. NSCs were cultured under four different conditions: on flat cover slips (2-D structure) in media with or without NGF, and on chitosan scaffold (3-D structure) in media with or without NGF. Immunohistochemical staining was used to observe multi-directional differentiation of cultured NSCs. Photomicrographs were taken and analyzed for cell number, soma size, and neuronal process length.
The porosity index of chitosan scaffold was around 90% and the diameter of pores was 50-350 μm. NSCs could differentiate into neurons, astrocytes, and oligodendrocytes under all culture conditions. The rank efficacy for neuronal differentiation was 3-D culture with NGF group > 3-D culture without NGF group > 2-D culture with NGF group > 2-D culture without NGF group.
The results suggest that the combination of chitosan scaffold and NGF exerts a synergistic effect on neuronal differentiation of NSCs, a requirement for successful integration into the damaged central nervous system.
成功的神经干细胞（NSCs）治疗需要将NSCs可控地分化为神经元。研究了在三维（3-D）培养中，多孔壳聚糖支架在神经生长因子（NGF）存在的情况下是否能促进NSCs向神经元分化。
采用冷冻干燥技术制备壳聚糖支架。NSCs在四种不同条件下培养：在有无NGF的培养基中于平的盖玻片上（二维结构），以及在有无NGF的培养基中于壳聚糖支架上（三维结构）。免疫组织化学染色用于观察培养的NSCs的多向分化。拍摄显微照片并分析细胞数量、细胞体大小和神经突长度。
壳聚糖支架的孔隙率约为90%，孔径为50 - 350μm。在所有培养条件下，NSCs均可分化为神经元、星形胶质细胞和少突胶质细胞。神经元分化的效果排序为：含NGF的三维培养组＞不含NGF的三维培养组＞含NGF的二维培养组＞不含NGF的二维培养组。
结果表明，壳聚糖支架与NGF的组合对NSCs向神经元的分化具有协同作用，这是成功整合到受损中枢神经系统的必要条件。
