Khaswaneh Ramada R, Abu-El-Rub Ejlal, Alzu'bi Ayman, Almazari Rawan, Alrabie Amneh, Almahasneh Fatimah A, Kasasbeh Amani, Ai-Jariri Heba F, Mustafa Ayman
Department of Basic Medical Sciences, Faculty of Medicine, Yarmouk University, Irbid, Jordan.
Department of Basic Medical Sciences, College of Medicine, QU Health, Qatar University, Doha, Qatar.
J Neuropathol Exp Neurol. 2025 Aug 1;84(8):680-691. doi: 10.1093/jnen/nlaf025.
Recent research indicates that mesenchymal stem cells (MSCs) can transdifferentiate into neuron-like cells under specific conditions, offering promise for neuronal regeneration. However, challenges remain in optimizing differentiation protocols to generate specific neuron types. This study explores the impact of supplementing neuronal induction media with dopamine and vitamin E to guide MSCs toward specific neuronal subtypes. Human adipose-MSCs were utilized to investigate neuronal differentiation. The cells were cultured in induction media supplemented with 2 concentrations of dopamine (2.5 and 5 µM) and vitamin E (12.5 and 25 µM). Immunostaining and western blot analysis were employed to assess the sequential expression of neuronal markers associated with various stages of maturation and development. These markers included Nestin, MAP2, NeuN, TBR1, SATB2, DAT, DBH, and CHAT. The results demonstrate, for the first time, that supplementing neuronal induction media with dopamine and vitamin E significantly enhances and accelerates the differentiation of MSCs into neuronal cells. Furthermore, the findings suggest that the induced cells are predominantly reprogrammed toward a cholinergic neuronal lineage. For MSCs, our study reveals that the addition of dopamine and vitamin E reprograms MSCs mainly toward cholinergic neurons, suggesting promising approaches for treating neurodegenerative disorders.
近期研究表明，间充质干细胞（MSCs）在特定条件下可转分化为神经元样细胞，为神经元再生带来了希望。然而，在优化分化方案以生成特定神经元类型方面仍存在挑战。本研究探讨了在神经元诱导培养基中添加多巴胺和维生素E对引导MSCs向特定神经元亚型分化的影响。利用人脂肪来源的MSCs研究神经元分化。将细胞培养在添加了2种浓度多巴胺（2.5和5 μM）和维生素E（12.5和25 μM）的诱导培养基中。采用免疫染色和蛋白质印迹分析来评估与成熟和发育各阶段相关的神经元标志物的顺序表达。这些标志物包括巢蛋白、微管相关蛋白2（MAP2）、神经元核抗原（NeuN）、T-框蛋白1（TBR1）、特异AT序列结合蛋白2（SATB2）、多巴胺转运体（DAT）、多巴胺β-羟化酶（DBH）和胆碱乙酰转移酶（CHAT）。结果首次表明，在神经元诱导培养基中添加多巴胺和维生素E可显著增强并加速MSCs向神经元细胞的分化。此外，研究结果表明，诱导细胞主要重编程为胆碱能神经元谱系。对于MSCs而言，我们的研究表明，添加多巴胺和维生素E可使MSCs主要重编程为胆碱能神经元，为治疗神经退行性疾病提供了有前景的方法。
