Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Department of Cell and Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran.
Hum Cell. 2021 Jul;34(4):1142-1152. doi: 10.1007/s13577-021-00537-9. Epub 2021 Apr 25.
Over the past decades, stem cell therapy has been investigated as a promising approach towards various diseases, including neurodegenerative disorders. Stem cells show the capability to differentiate into neuronal progenitor cells in vitro. In the present study, the differentiation potential of human-induced pluripotent stem cells (hiPSCs) into neural lineages was examined under the efficient induction media containing forskolin and 3-isobutyl-1-methyl-xanthine (IBMX) in the presence of nisin (Ni), non-essential amino acids (NEAA) and combination of those (NEAA-Ni) in vitro. The optimum concentrations of these factors were obtained by MTT assay and acridine orange (AO) staining. The effect of Ni and NEAA on the expression rate of neural-specific markers including NSE, MAP2, and ß-tubulin III was studied via immunocytochemistry (ICC) and real-time RT-PCR analyses. Our results indicated that the induction medium containing Ni or NEAA increased the gene and protein expression of NSE, MAP2, and β-tubulin III on the 14th differentiation day. On the other hand, NEAA-Ni showed a less-differentiated hiPSCs compared to Ni and NEAA alone. In conclusion, the obtained results illustrated that Ni and NEAA could be applied as effective factors for neural differentiation of hiPSCs in the future.
在过去的几十年中，干细胞治疗作为一种有前途的方法，已经被广泛研究用于治疗各种疾病，包括神经退行性疾病。干细胞具有在体外分化为神经元祖细胞的能力。在本研究中，研究了在含有佛司可林和 3-异丁基-1-甲基黄嘌呤（IBMX）的有效诱导培养基中，人诱导多能干细胞（hiPSCs）在存在乳链菌肽（Ni）、非必需氨基酸（NEAA）和它们的组合（NEAA-Ni）的情况下向神经谱系分化的潜力。通过 MTT 测定和吖啶橙（AO）染色获得了这些因素的最佳浓度。通过免疫细胞化学（ICC）和实时 RT-PCR 分析研究了 Ni 和 NEAA 对神经特异性标志物包括 NSE、MAP2 和β-微管蛋白 III 的表达率的影响。我们的结果表明，含有 Ni 或 NEAA 的诱导培养基在第 14 天分化时增加了 NSE、MAP2 和β-微管蛋白 III 的基因和蛋白表达。另一方面，与 Ni 和 NEAA 单独使用相比，NEAA-Ni 显示出分化程度较低的 hiPSCs。总之，研究结果表明，Ni 和 NEAA 可以作为未来 hiPSCs 神经分化的有效因素。
