Abdullah Rafal H, Yaseen Nahi Y, Salih Shahlaa M, Al-Juboory Ahmad Adnan, Hassan Ayman, Al-Shammari Ahmed Majeed
Experimental Therapy Department, Iraqi Center for Cancer and Medical Genetic Research, Al-Mustansiriyah University, Baghdad, Iraq.
Department of Biotechnology, Al-Nahrain University, Baghdad, Iraq.
J Chem Neuroanat. 2016 Nov;77:129-142. doi: 10.1016/j.jchemneu.2016.07.003. Epub 2016 Jul 11.
The differentiation of mesenchymal stem cells (MSC) into acetylcholine secreted motor neuron-like cells, followed by elongation of the cell axon, is a promising treatment for spinal cord injury and motor neuron cell dysfunction in mammals. Differentiation is induced through a pre-induction step using Beta- mercaptoethanol (BME) followed by four days of induction with retinoic acid and sonic hedgehog. This process results in a very efficient differentiation of BM-MSCs into motor neuron-like cells. Immunocytochemistry showed that these treated cells had specific motor neural markers: microtubule associated protein-2 and acetylcholine transferase. The ability of these cells to function as motor neuron cells was assessed by measuring acetylcholine levels in a culture media during differentiation. High-performance liquid chromatography (HPLC) showed that the differentiated cells were functional. Motor neuron axon elongation was then induced by adding different concentrations of a nerve growth factor (NGF) to the differentiation media. Using a collagen matrix to mimic the natural condition of neural cells in a three-dimensional model showed that the MSCs were successfully differentiated into motor neuron-like cells. This process can efficiently differentiate MSCs into functional motor neurons that can be used for autologous nervous system therapy and especially for treating spinal cord injuries.
间充质干细胞（MSC）分化为分泌乙酰胆碱的运动神经元样细胞，随后细胞轴突伸长，这是一种治疗哺乳动物脊髓损伤和运动神经元细胞功能障碍的有前景的方法。分化通过使用β-巯基乙醇（BME）进行预诱导步骤，然后用视黄酸和音猬因子进行四天诱导来实现。该过程导致骨髓间充质干细胞（BM-MSCs）非常有效地分化为运动神经元样细胞。免疫细胞化学显示，这些处理过的细胞具有特定的运动神经标记物：微管相关蛋白-2和乙酰胆碱转移酶。通过在分化过程中测量培养基中的乙酰胆碱水平来评估这些细胞作为运动神经元细胞的功能。高效液相色谱（HPLC）显示分化后的细胞具有功能。然后通过向分化培养基中添加不同浓度的神经生长因子（NGF）来诱导运动神经元轴突伸长。在三维模型中使用胶原基质模拟神经细胞的自然条件表明，间充质干细胞成功分化为运动神经元样细胞。这一过程可以有效地将间充质干细胞分化为功能性运动神经元，可用于自体神经系统治疗，特别是用于治疗脊髓损伤。
