新生小鼠神经干细胞的分离、培养和鉴定

目的：探讨新生小鼠神经干细胞(NSCs)分离、培养以及鉴定的方法。方法：分离新生昆明种小鼠(出生24h内)的大脑组织,经胰酶消化加机械吹打后,利用无血清培养基悬浮培养细胞,获得具有自我增殖能力的细胞克隆,并将细胞克隆贴壁培养测其分化能力。应用抗Ncstin、Musashi、SSEA-1、NSE免疫细胞化学方法及BrdU标记方法鉴定细胞克隆。结果：从新生昆明种小鼠大脑组织分离的细胞悬液,经悬浮培养,生成大量具有增殖能力的细胞,可形成神经球(neurosphercs),抗Nestin、Musashi、SSEA-1阳性,BrdU标记也呈阳性。细胞克隆贴壁培养后神经球分化为神经细胞,抗NSE阳性。结论：上述方法分离的细胞具有自我更新、自我复制及分化为神经细胞的能力,属于中枢神经系统干细胞。     

Directing neural stem cell fate with biomaterial parameters for injured brain regeneration

The discovery of neural stem cells (NSCs), which have the ability to self-renew and differentiate into all types of neural lineages, offers promising prospect for the treatment of brain neurological disorders such as stroke/cerebral ischemia, traumatic brain injury and neurodegenerative disorders. However, only limited number of NSCs could survive or propagate due to tissue inflammation or blood-brain barrier. Therefore, it is necessary to develop an appropriate culture system that highly mimics the natural NSCs niche to direct stem cell fate and behavior for nerve regeneration. Both biophysical and biochemical properties of the NSC niche, including topology, mechanical properties, bioactive molecules, and their spatial and temporal presentations should be considered for the design of functionalized scaffolds, which could not only serve as the delivery vehicles of NSCs but also stimulate specific cellular responses at the molecular level, such as support endogenous or exogenous cells proliferation, migration and homing, even promote the growth of axon at the injured brain site. This review attempts to outline the varieties of biomaterial     

神经球内神经干/祖细胞活性与代谢研究

为定量研究静态无血清悬浮培养的神经球内神经干/祖细胞的活性及代谢情况,取孕14 d的小鼠胚胎,从海马脑组织分离出神经干/祖细胞,培养过程中跟踪测定神经球直径的分布变化,利用CCK-8法测定活细胞数,同时对培养上清进行葡萄糖、乳酸、谷氨酰胺和氨的定量测定,并对相关细胞代谢动力学参数进行计算和分析.结果表明在培养过程中,神经球内细胞活性及细胞代谢均随着球直径的增加而减弱.在神经球平均直径为100 μm左右,培养基中葡萄糖、谷氨酰胺浓度分别为36.38 mmol/L和1.33 mmol/L时,球心细胞生长开始受到抑制;而当神经球直径主体分布于100～150 μm,培养基中葡萄糖浓度为31.11 mmol/L、谷氨酰胺浓度为1.15 mmol/L时,神经球内部细胞死亡数开始大于增殖数,同时细胞代谢降低到很低水平.表明随着神经球直径的增加而造成的传质困难对于其内部细胞的活性和代谢有重要影响.