毛囊神经嵴干细胞源性运动神经元、胶质细胞联合移植治疗肌萎缩脊髓侧索硬化的应用基础研究

The amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease and has not effective treatment so far. Although positive results of stem cells transplantation have been shown in various recent researches, the therapeutic effect of this approach is insufficient to meet the clinical requirement. Based on the new understanding of pathogenesis of ALS that degeneration and loss of motoneurons are key pathogenic events, and glial cells (main component of microenvironment) contribute to the initiation and improvement of ALS, a hypothesis was proposed that protection and supplement of motoneuron simultaneously performed should be benefit to cure ALS. In present experimental plan, the mice (SOD1G93A) were used to duplicate the ALS model, then, the motoneurones and glial cells derived from allogenic hair follicle neural crest stem cells were transplanted into the ventral horn of spinal cord. The transplanted motoneurons were intended to add and/or replace the degenerative and/or lost motoneurons, while the transplanted glial cells were intended to improve the microenvironment of spinal cord in mice with ALS, modulate and improve function of activated and overgrew glial cells, in order to protect the motoneurons and promote regeneration of motoneuron. The therapeutic effect of cotransplantation of stem cells derived motoneurons and glial cells was evaluated and mechanism underlying this effect was explored in this study, so, the data from this study may provide experimental basis for stem cells therapy of ALS.

ALS是一种致死性神经系统变性疾病，迄今尚缺乏有效的治疗手段。近年来，干细胞治疗ALS的研究展示了一定的治疗效果，但离人们的期望尚远。我们基于对运动神经元变性、丢失的致病关键，和胶质细胞微环境在运动神经元病变中的始动、促进作用的，ALS发病机制的新认识，提出了“利用干细胞移植对ALS运动神经元保护与补充同步进行”的设想。本课题拟以SOD1G93A基因突变小鼠为实验模型，将同种异体毛囊神经嵴干细胞源性运动神经元细胞和胶质细胞移植至脊髓灰质前角，意在一方面利用移植的运动神经元补充、替代变性、丢失的运动神经元细胞；另一方面利用移植的正常神经胶质细胞，改善ALS脊髓微环境，调控和改善活化增生的胶质细胞的功能、保护运动神经元、促进运动神经元细胞的再生；旨在通过干细胞源性运动神经元细胞与胶质细胞联合移植对ALS治疗的效果观测及机制探讨，为临床治疗ALS提供一种新的治疗方案。

肌萎缩脊髓侧索硬化症（ Amyotrophic lateralsclerosis ，ALS）是一种致死性神经系统变性疾病，迄今尚缺乏有效的治疗手段。我们基于对运动神经元变性、丢失的致病关键，和胶质细胞微环境在运动神经元病变中的始动、促进作用的ALS发病机制新认识，提出了“利用干细胞移植对ALS运动神经元保护与补充同步进行”的设想。本课题以SOD1G93A基因突变小鼠为模型，将同种异体毛囊神经嵴干细胞源性运动神经元细胞和胶质细胞联合移植至脊髓灰质前角，观察干细胞联合移植对ALS的治疗作用及机制。得出以下主要结论：.1、毛囊神经嵴干细胞（（hair follicle neural crest stem cell，hfNCSCs）可来源自体、取材方便、易于诱导分化、可自然分化为神经元和神经胶质细胞，是神经干细胞移植较为理想的种子细胞。.2、利用转录因子、细胞因子、小分子物质等非基因干涉的方法，经向运动神经元前体细胞、运动神经元二步的诱导分化方法，可将hfNCSCs诱导分化为具有运动神经元结构、电生理特性和功能的运动神经元细胞。.3、ScienCell星形胶质细胞培养基，可将hfNCSCs诱导分化为星形胶质细胞。.4、体外实验证实，hfNCSCs源性星形胶质细胞可以降低运动神经元的凋亡速度，促进运动神经元的增殖。.5、ALS小鼠脊髓灰质前角，同种异体hfNCSCs源性运动神经元、星形胶质细胞联合移植，移植细胞可以存活、迁移，减缓脊髓灰质前角运动神经元和移植的干细胞源性运动神经元细胞的变性凋亡，维持骨骼肌的神经支配，延长ALS小鼠生存期。. 本研究结果初步证明，通过移植正常星形胶质细胞改善局部微环境的方法，可以减缓脊髓灰质前角运动神经元和移植运动神经元的变性，延缓ALS病程进展， 发挥治疗ALS作用，值得j进行深入研究。