Singh Varsha, Sherpa Mingma
Chitkara School of Health Sciences, Chitkara University, Patiala, Punjab, India.
Department of Pathology, Sikkim Manipal Institute of Medical Science, Gangtok, Sikkim, India.
Appl Biochem Biotechnol. 2017 Sep;183(1):385-395. doi: 10.1007/s12010-017-2452-1. Epub 2017 Mar 13.
Mesenchymal stem cells have been extensively used for cell-based therapies especially in neuronal diseases. Studies still continue to delineate mechanisms involved in differentiating mesenchymal stem cells into neuronal cells under experimental conditions as they have low mortality rate and hence, the number of cells available for experiments is much more limited. Culturing and differentiating of neuronal cell is more challenging as they do not undergo cell division thus, bringing them to differentiate proves to be a difficult task. Here, the aim of this study is to investigate whether Juglans regia L. (walnut oil) differentiates multipotent, C3H10T1/2 cells, a murine mesenchymal stem cell line, into neuronal cells. A simple treatment protocol induced C3H10T1/2 cells to exhibit a neuronal phenotype. With this optimal differentiation protocol, almost all cells exhibited neuronal morphology. The cell bodies extended long processes. C3H10T1/2 cells were plated and treated with walnut oil post 24 h of plating. The treatment was given (with walnut oil treated cultures with or without control cultures) at different concentrations. The cultured cells were then stained with cresyl violet acetate solution which was used to stain the Nissl substance in the cytoplasm of the induced neuronal culture. The results indicated that the C3H10T1/2 cells differentiated into neuronal-like cells with long outgrowths of axon-like structures able to take up the cresyl violet acetate stain indicating their preliminary differentiation into neuronal-like morphology with walnut oil treatment. Treating the mesenchymal stem cells can in future establish a cultured mesenchymal stem cell line as neuronal differentiating cell line model.
间充质干细胞已被广泛用于基于细胞的治疗，尤其是在神经疾病方面。由于间充质干细胞死亡率低，因此在实验条件下可用于实验的细胞数量更为有限，相关研究仍在继续阐明将间充质干细胞分化为神经细胞所涉及的机制。培养和分化神经细胞更具挑战性，因为它们不会进行细胞分裂，因此，促使它们分化是一项艰巨的任务。在此，本研究的目的是调查胡桃（核桃油）是否能将多能性的小鼠间充质干细胞系C3H10T1/2细胞分化为神经细胞。一个简单的处理方案诱导C3H10T1/2细胞呈现神经表型。通过这种优化的分化方案，几乎所有细胞都呈现出神经形态。细胞体伸出长长的突起。接种C3H10T1/2细胞，接种24小时后用核桃油处理。以不同浓度进行处理（处理核桃油处理的培养物，有无对照培养物）。然后用醋酸甲酚紫溶液对培养的细胞进行染色，该溶液用于对诱导的神经培养物细胞质中的尼氏体进行染色。结果表明，C3H10T1/2细胞分化为具有轴突样结构长突起的神经样细胞，能够摄取醋酸甲酚紫染色，表明经核桃油处理后它们初步分化为神经样形态。未来，处理间充质干细胞可以建立一个培养的间充质干细胞系作为神经分化细胞系模型。
