Mesenchymal stem cells (MSCs) isolated from bone marrow have different developmental origins, including neural crest. MSCs can differentiate into neural progenitor-like cells (NPCs) under the influence of bFGF and EGF. NPCs can terminally differentiate into neurons that express beta-III-tubulin and elicit action potential. The main aim of the study was to identify key genetic markers involved in differentiation of MSCs into NPCs through transcriptomic analysis. Total RNA was isolated from MSCs and MSCs-derived NPCs followed by cDNA library construction for transcriptomic analysis. Sample libraries that passed the quality and quantity assessments were subjected to high throughput mRNA sequencing using NextSeq®500. Differential gene expression analysis was performed using the DESeq2 R package with MSC samples being a reference group. The expression of eight differentially regulated genes was counter validated using real-time PCR. In total, of the 3,252 differentially regulated genes between MSCs and NPCs with two or more folds, 1,771 were upregulated genes, whereas 1,481 were downregulated in NPCs. Amongst these differential genes, 104 transcription factors were upregulated, and 45 were downregulated in NPCs. Neurogenesis related genes were upregulated in NPCs and the main non-redundant gene ontology (GO) terms enriched in NPCs were the autonomic nervous system, cell surface receptor signalling pathways), extracellular structure organisation, and programmed cell death. The main non-redundant GO terms enriched in MSCs included cytoskeleton organisation cytoskeleton structural constituent, mitotic cell cycle), and the mitotic cell cycle process Gene set enrichment analysis also confirmed cell cycle regulated pathways as well as Biocarta integrin pathway were upregulated in MSCs. Transcription factors enrichment analysis by ChEA3 revealed Foxs1 and HEYL, amongst the top five transcription factors, inhibits and enhances, respectively, the NPCs differentiation of MSCs. The vast differences in the transcriptomic profiles between NPCs and MSCs revealed a set of markers that can identify the differentiation stage of NPCs as well as provide new targets to enhance MSCs differentiation into NPCs.

中文翻译：

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显着的转录组变化与 bFGF 和 EGF 存在下骨髓来源的间充质干细胞分化为神经祖细胞样细胞有关

从骨髓中分离的间充质干细胞（MSC）具有不同的发育起源，包括神经嵴。MSCs在bFGF和EGF的影响下可以分化为神经祖细胞样细胞（NPCs）。NPC 最终可以分化为表达 β-III-微管蛋白并引发动作电位的神经元。该研究的主要目的是通过转录组分析鉴定参与 MSC 分化为 NPC 的关键遗传标记。从 MSC 和 MSC 衍生的 NPC 中分离总 RNA，然后构建 cDNA 文库用于转录组分析。通过质量和数量评估的样品文库使用 NextSeq®500 进行高通量 mRNA 测序。使用DESeq2 R软件包以MSC样本作为参考组进行差异基因表达分析。使用实时 PCR 对八个差异调节基因的表达进行了反验证。总共，在 MSC 和 NPC 之间具有两个或更多折叠的 3,252 个差异调节基因中，1,771 个上调基因，而 NPC 中有 1,481 个下调基因。在这些差异基因中，104个转录因子在NPC中上调，45个转录因子下调。神经发生相关基因在NPC中上调，NPC中富集的主要非冗余基因本体（GO）术语是自主神经系统、细胞表面受体信号通路、细胞外结构组织和程序性细胞死亡。MSCs中富集的主要非冗余GO术语包括细胞骨架组织（细胞骨架结构成分、有丝分裂细胞周期）、有丝分裂细胞周期过程基因集富集分析也证实细胞周期调节通路以及Biocarta整合素通路在MSCs中上调。ChEA3 的转录因子富集分析显示，排名前 5 位的转录因子中的 Foxs1 和 HEYL 分别抑制和增强 MSC 的 NPC 分化。NPC 和 MSC 之间转录组图谱的巨大差异揭示了一组标记，可以识别 NPC 的分化阶段，并提供增强 MSC 向 NPC 分化的新靶标。