METHODS: The human ES cell line H1 was used for the experiments. hESCs were treated with an acoustic vibration device. Their proliferative ability was subsequently detected using a colony formation assay, while the expression of pluripotency-related markers was detected via immunofluorescence staining. Finally, changes in gene expression levels were examined using quantitative polymerase chain reaction (qPCR) in the presence of appropriate primers.
RESULTS: Compared with normal cells in the control group, the morphology of experimental cells subjected to acoustic vibration did not significantly change. Contrastingly, the colony-forming efficiency of the experimental cells significantly increased. Immunofluorescence staining results showed the cells in experimental group were positive for the pluripotency markers NANOG, octamer-binding transcription factor 4 gene (OCT4), and SRY (sex determining region Y)-box 2 (SOX2). In addition, the expression levels of pluripotency genes NANOG, OCT4, SOX2, and Yes-associated protein (YAP)-related genes were up-regulated following acoustic vibration.
CONCLUSIONS: Our results revealed that acoustic vibration enhanced the proliferative ability of hESCs and increased the expression levels of NANOG, OCT4, SOX2, and YAP-related genes, indicating that acoustic vibration can optimize the self-renewal ability of hESCs and that the YAP signaling pathway may play a critical role in the functional process of acoustic vibration.
方法:实验使用人ES细胞系H1。用声振动装置处理hESC。随后使用集落形成测定法检测它们的增殖能力,而通过免疫荧光染色检测到多能性相关标志物的表达。最后,在适当的引物存在下,使用定量聚合酶链反应(qPCR)检测基因表达水平的变化.
结果:与对照组的正常细胞相比,受声振动作用的实验细胞形态无明显变化。相反,实验细胞的集落形成效率显着提高。免疫荧光染色结果显示实验组细胞多能性标志物NANOG阳性,八聚体结合转录因子4基因(OCT4),和SRY(性别决定区Y)-框2(SOX2)。此外,多能性基因NANOG的表达水平,OCT4,SOX2和Yes相关蛋白(YAP)相关基因在声振动后上调。
结论:我们的结果表明,声振动增强了hESCs的增殖能力,并增加了NANOG的表达水平,OCT4、SOX2和YAP相关基因,表明声振动可以优化hESCs的自我更新能力,YAP信号通路可能在声振动的功能过程中起关键作用。