Preparation of Neural Stem Cells and Progenitors: Neuronal Production and Grafting Applications
Preparation of Neural Stem Cells and Progenitors: Neuronal Production and Grafting Applications
复制标题
DOI：
10.1007/978-1-62703-640-5_7
复制
发表时间：
2013-01-01
期刊：
影响因子：
--
通讯作者：
Fischer, Itzhak
中科院分区：
文献类型：
--
作者：
Bonner, Joseph F.;Haas, Christopher J.;Fischer, Itzhak
神经干细胞（NSC）不仅是研究神经发育和功能的重要工具，也是神经疾病移植策略发展的重要组成部分。NSC可以用来研究神经元如何获得不同的表型，以及神经系统发育中神经元和神经胶质之间的相互作用如何塑造中枢神经系统（CNS）的结构和功能。此外，由NSC制备的神经元可用于阐明神经系统疾病的分子基础以及潜在的治疗方法。虽然NSC可以从不同的物种和许多来源获得，包括胚胎干细胞、诱导多能干细胞、成体中枢神经系统和非神经细胞的直接重编程，但直接从大鼠胎儿组织中分离原代NSC是制备和研究神经元最常用的技术，具有丰富的可用于比较的数据。无论来源材料如何，类似的技术都被用于在培养中维持NSC，并将NSC分化为成熟的神经谱系。本章将描述从胚胎大鼠中枢神经系统组织（主要是脊髓）中分离多能NSC和神经前体细胞（NPC）的具体方法。特别是，鼻咽癌可以分离成神经元和胶质限制性前体（分别为NRP和GRP），并在体外和移植到成人中枢神经系统后可靠地产生神经元或胶质细胞。本章将详细描述从大鼠脊髓中分离出的NSC和NPC用于随后的体外或体内研究所需的分离、繁殖、储存和分化方法。
Neural stem cells (NSC) are not only a valuable tool for the study of neural development and function, but an integral component in the development of transplantation strategies for neural disease. NSC can be used to study how neurons acquire distinct phenotypes and how the reciprocal interactions between neurons and glia in the developing nervous system shape the structure and function of the central nervous system (CNS). In addition, neurons prepared from NSC can be used to elucidate the molecular basis of neurological disorders as well as potential treatments. Although NSC can be derived from different species and many sources, including embryonic stem cells, induced pluripotent stem cells, adult CNS, and direct reprogramming of non-neural cells, isolating primary NSC directly from rat fetal tissue is the most common technique for preparation and study of neurons with a wealth of data available for comparison. Regardless of the source material, similar techniques are used to maintain NSC in culture and to differentiate NSC toward mature neural lineages. This chapter will describe specific methods for isolating multipotent NSC and neural precursor cells (NPC) from embryonic rat CNS tissue (mostly spinal cord). In particular, NPC can be separated into neuronal and glial restricted precursors (NRP and GRP, respectively) and used to reliably produce neurons or glial cells both in vitro and following transplantation into the adult CNS. This chapter will describe in detail the methods required for the isolation, propagation, storage, and differentiation of NSC and NPC isolated from rat spinal cords for subsequent in vitro or in vivo studies.