Montoya-Gacharna Jose V, Sutachan Jhon Jairo, Chan Wai Si, Sideris Alexandra, Blanck Thomas J J, Recio-Pinto Esperanza
Department of Anesthesiology, Langone Medical Center, New York University, New York, NY, USA.
Methods Mol Biol. 2012;846:103-16. doi: 10.1007/978-1-61779-536-7_10.
Spinal cord motor neuron cultures are an important tool for the study of mechanisms involved in motor neuron survival, degeneration and regeneration, volatile anesthetic-induced immobility, motor neuron disorders such as amyotrophic lateral sclerosis or spinal muscular atrophy as well as in spinal cord injury. Embryonic spinal cord motor neurons derived from rats have been successfully cultured; unfortunately, the culture of adult motor neurons has been problematic due to their short-term survival. Recently, by using a cocktail of target-derived factors, neurotrophins (brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor) and a permeable cyclic adenosine monophosphate analog, we have established a reproducible protocol for long-term cultures of healthy and functional adult motor neurons (Exp Neurol 220:303-315, 2009). Here, we now describe in detail the steps that we used for the optimization of the process of isolation and maintenance of adult rat ventral horn motor neurons in vitro.
脊髓运动神经元培养是研究运动神经元存活、退化和再生机制、挥发性麻醉剂诱导的不动、运动神经元疾病（如肌萎缩侧索硬化症或脊髓性肌萎缩症）以及脊髓损伤的重要工具。源自大鼠的胚胎脊髓运动神经元已成功培养；不幸的是，由于成年运动神经元存活时间短，其培养一直存在问题。最近，通过使用目标衍生因子、神经营养因子（脑源性神经营养因子和胶质细胞系衍生的神经营养因子）和一种可渗透的环磷酸腺苷类似物的混合物组合，我们建立了一种可重复的方案，用于健康且功能正常的成年运动神经元的长期培养（《实验神经病学》220:303 - 315，2009年）。在此，我们现在详细描述用于优化体外分离和维持成年大鼠腹角运动神经元过程的步骤。
