Döbrössy Máté, Pruszak Jan
University Medical Center Freiburg, Freiburg, Germany.
Methods Mol Biol. 2013;1037:117-44. doi: 10.1007/978-1-62703-505-7_7.
The nervous system is characterized by its complex network of highly specialized cells that enable us to perceive stimuli from the outside world and react accordingly. The computational integration enabled by these networks remains to be elucidated, but appropriate sensory input, processing, and motor control are certainly essential for survival. Consequently, loss of nervous tissue due to injury or disease represents a considerable biomedical challenge. Stem cell research offers the promise to provide cells for nervous system repair to replace lost and damaged neural tissue and alleviate disease. We provide a protocol-based chapter on fundamental principles and procedures of pluripotent stem cell (PSC) differentiation and neural transplantation. Rather than detailed methodological step-by-step descriptions of these procedures, we provide an overview and highlight the most critical aspects and key steps of PSC neural induction, subtype specification in different in vitro systems, as well as neural cell transplantation to the central nervous system. We conclude with a summary of suitable readout methods including in vitro phenotypic analysis, histology, and functional analysis in vivo.
神经系统的特征在于其由高度专业化细胞构成的复杂网络，这些细胞使我们能够感知来自外部世界的刺激并做出相应反应。这些网络实现的计算整合仍有待阐明，但适当的感觉输入、处理和运动控制对于生存肯定至关重要。因此，由于损伤或疾病导致的神经组织损失是一项重大的生物医学挑战。干细胞研究有望提供用于神经系统修复的细胞，以替代丢失和受损的神经组织并缓解疾病。我们提供了一章基于方案的内容，介绍多能干细胞（PSC）分化和神经移植的基本原理及程序。我们并非对这些程序进行详细的方法步骤描述，而是提供一个概述，并突出PSC神经诱导、不同体外系统中的亚型特异性以及向中枢神经系统进行神经细胞移植的最关键方面和关键步骤。我们最后总结了合适的读出方法，包括体外表型分析、组织学和体内功能分析。
