Majeed Saba, Aziz Aisha, Simjee Shabana Usman
H.E.J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Pakistan.
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Pakistan.
Heliyon. 2020 Oct 8;6(10):e05083. doi: 10.1016/j.heliyon.2020.e05083. eCollection 2020 Oct.
Several approaches have been applied to harvest bone marrow stromal cells (BMSCs) and to differentiate into neurons or neuronal-like cells through chemical stimulation or exposing to growth factors. To date, the data regarding induction or regulation of neuronal transcription program in neuronal-like cells derived from BMSCs is yet unknown. The objective of this study is to co-culture BMSCs with neonatal hippocampal cells and generate neuronal-like cells by direct cell-to-cell contact system without using neuronal growth factors or neurobasal medium. Here, we proposed a role for NeuroD1 and Neurogenin -2 bHLH family of transcription factors implicated in onset of neurogenesis and differentiation of cells into neurons in promoting the interaction of hippocampal cells with BMSCs and their differentiation in to neurons. The proliferation of the cells was assessed with MTT assay and the role of neuronal induction and differentiation transcription regulators NeuroD1 and Neurogenin-2 in cocultured cells was determined through immunocytochemical analysis. We observed activation and expression of the neurogenic transcription factors NeuroD1 and NGN-2 associated with neuronal activation program to initiate the onset of neurogenesis in cocultured cells. Further, our results have shown a significant expression of neuronal progenitor and immature neuronal marker i.e., nestin and tubulin respectively in cocultured cells endorsing the initiation of neuronal activation.
已经应用了几种方法来获取骨髓基质细胞（BMSC），并通过化学刺激或暴露于生长因子使其分化为神经元或神经元样细胞。迄今为止，关于骨髓间充质干细胞来源的神经元样细胞中神经元转录程序的诱导或调节的数据尚不清楚。本研究的目的是将骨髓间充质干细胞与新生海马细胞共培养，并通过直接细胞间接触系统生成神经元样细胞，而不使用神经元生长因子或神经基础培养基。在此，我们提出了NeuroD1和Neurogenin-2 bHLH转录因子家族在神经发生起始和细胞向神经元分化中的作用，它们参与促进海马细胞与骨髓间充质干细胞的相互作用及其向神经元的分化。用MTT法评估细胞增殖，并通过免疫细胞化学分析确定神经元诱导和分化转录调节因子NeuroD1和Neurogenin-2在共培养细胞中的作用。我们观察到与神经元激活程序相关的神经源性转录因子NeuroD1和NGN-2的激活和表达，从而启动共培养细胞中神经发生的起始。此外，我们的结果显示，共培养细胞中分别显著表达神经元祖细胞和未成熟神经元标志物，即巢蛋白和微管蛋白，这支持了神经元激活的起始。
