Xia Bin, Zou Yang, Xu Zhiling, Lv Yonggang
Key Laboratory of Biorheological Science and Technology (Chongqing University), Ministry of Education, Bioengineering College, Chongqing University, Chongqing, People's Republic of China.
Mechanobiology and Regenerative Medicine Laboratory, Bioengineering College, Chongqing University, Chongqing, People's Republic of China.
Biotechnol Appl Biochem. 2017 Nov;64(6):927-937. doi: 10.1002/bab.1554. Epub 2017 May 30.
Low-intensity pulsed ultrasound (LIPUS) is a noninvasive technique that has been shown to affect cell proliferation, migration, and differentiation and promote the regeneration of damaged peripheral nerve. Our previous studies had proved that LIPUS can significantly promote the neural differentiation of induced pluripotent stem cell-derived neural crest stem cells (iPSCs-NCSCs) and enhance the repair of rat-transected sciatic nerve. To further explore the underlying mechanisms of LIPUS treatment of iPSCs-NCSCs, this study reported the gene expression profiling analysis of iPSCs-NCSCs before and after LIPUS treatment using the RNA-sequencing (RNA-Seq) method. It was found that expression of 76 genes of iPSCs-NCSCs cultured in a serum-free neural induction medium and expression of 21 genes of iPSCs-NCSCs cultured in a neuronal differentiation medium were significantly changed by LIPUS treatment. The differentially expressed genes are related to angiogenesis, nervous system activity and functions, cell activities, and so on. The RNA-seq results were further verified by a quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR). High correlation was observed between the results obtained from qRT-PCR and RNA-Seq. This study presented new information on the global gene expression patterns of iPSCs-NCSCs after LIPUS treatment and may expand the understanding of the complex molecular mechanism of LIPUS treatment of iPSCs-NCSCs.
低强度脉冲超声(LIPUS)是一种非侵入性技术,已被证明可影响细胞增殖、迁移和分化,并促进受损周围神经的再生。我们之前的研究已经证明,LIPUS可以显著促进诱导多能干细胞来源的神经嵴干细胞(iPSCs-NCSCs)的神经分化,并增强大鼠坐骨神经横断后的修复。为了进一步探索LIPUS治疗iPSCs-NCSCs的潜在机制,本研究报告了使用RNA测序(RNA-Seq)方法对LIPUS治疗前后iPSCs-NCSCs进行基因表达谱分析的结果。研究发现,在无血清神经诱导培养基中培养的iPSCs-NCSCs的76个基因的表达以及在神经元分化培养基中培养的iPSCs-NCSCs的21个基因的表达在LIPUS处理后发生了显著变化。差异表达基因与血管生成、神经系统活性和功能、细胞活性等有关。RNA测序结果通过定量实时逆转录聚合酶链反应(qRT-PCR)进一步验证。qRT-PCR结果与RNA-Seq结果之间观察到高度相关性。本研究提供了LIPUS处理后iPSCs-NCSCs全局基因表达模式的新信息,并可能扩展对LIPUS治疗iPSCs-NCSCs复杂分子机制的理解。