A stably established population of mouse bone marrow stromal cells (BMSCs) with self-renewal and multilineage differentiation potential was expanded in vitro for more than 50 passages. These cells express high levels of mesenchymal stem cell markers and can be differentiated into adipogenic, chondrogenic, and osteogenic lineages in vitro. Subjected to basic fibroblast growth factor (bFGF) treatment, a typical neuronal phenotype was induced in these cells, as supported by neuronal morphology, induction of neuronal markers, and relevant electrophysiological excitability. To identify the genes regulating neuronal differentiation, cDNA microarray analysis was conducted using mRNAs isolated from cells differentiated for different time periods (0, 4, 24, and 72 h) after bFGF treatment. Various expression patterns of neuronal genes were stimulated by bFGF. These gene profiles were shown to be involved in developmental, functional, and structural integration of the nervous system. The expression of representative genes stimulated by bFGF in each group was verified by RT-PCR. Amongst proneural genes, the mammalian achate-schute homolog 1 (Mash-1), a basic helix-loop-helix transcriptional factor, was further demonstrated to be significantly upregulated. Overexpression of Mash-1 in mouse BMSCs was shown to induce the expression of neuronal specific enolase (NSE) and terminal neuronal morphology, suggesting that Mash-1 plays an important role in the induction of neuronal differentiation of mouse BMSCs.

中文翻译：

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碱性成纤维细胞生长因子诱导的小鼠骨髓基质细胞神经元分化的早期基因谱

具有自我更新和多系分化潜能的稳定建立的小鼠骨髓基质细胞（BMSC）群体在体外扩增了超过50代。这些细胞表达高水平的间充质干细胞标志物，并可以在体外分化为成脂，成软骨和成骨谱系。接受碱性成纤维细胞生长因子（bFGF）处理后，在这些细胞中诱导了典型的神经元表型，并得到了神经元形态，神经元标记物的诱导和相关的电生理兴奋性的支持。为了鉴定调节神经元分化的基因，使用从bFGF治疗后不同时间段（0、4、24和72 h）分化的细胞中分离的mRNA进行cDNA微阵列分析。bFGF刺激神经元基因的各种表达方式。这些基因图谱显示与神经系统的发育，功能和结构整合有关。通过RT-PCR验证每组中bFGF刺激的代表性基因的表达。在proneural基因中，哺乳动物的achate-schute进一步证明，同源物1（Mash-1），一种基本的螺旋-环-螺旋转录因子，被显着上调。Mash-1在小鼠BMSCs中的过表达可诱导神经元特异性烯醇化酶（NSE）的表达和终末神经元形态，这表明Mash-1在诱导小鼠BMSCs神经元分化中起重要作用。