Nanosecond pulsed electric fields enhance mesenchymal stem cells differentiation via DNMT1-regulated OCT4/NANOG gene expression
Nanosecond pulsed electric fields enhance mesenchymal stem cells differentiation via DNMT1-regulated OCT4/NANOG gene expression
复制标题
纳秒脉冲电场通过 DNMT1 调节的 OCT4/NANOG 基因表达增强间充质干细胞分化
DOI：
10.1186/s13287-020-01821-5
复制
发表时间：
2020-07-22
影响因子：
7.5
通讯作者：
Ge, Zigang
中科院分区：
文献类型：
--
作者：
Li, Kejia;Ning, Tong;Ge, Zigang
背景
骨髓间充质干细胞（mesenchymal stem cells，MSCs）是组织形成和再生的基本细胞，其分化潜能的提高有多种策略。然而，这些策略相对低效，限制了应用。在本研究中，我们报道了一种新的有效的策略，纳秒脉冲电场（nsPEFs）刺激，它可以提高骨髓间充质干细胞的三系分化潜力，并进一步解释了背后的机制。
方法
通过组织学染色筛选出促进MSCs向三系分化的nsPEFs参数，并通过功能基因检测进一步证实nsPEFs的作用。为了探讨相应的机制，我们检测了多能性基因的表达及其启动子的甲基化状态。最后，我们靶向了受nsPEFs影响的DNA甲基转移酶。
结果
单纯预处理5个脉冲的nsPEFs刺激可显著促进骨髓间充质干细胞的体外三系分化（能量水平为10 ns，20 kV/cm; 100 ns，10 kV/cm），这是由于nsPEF通过瞬时下调DNA甲基化转移酶1（DNMT 1）使干细胞多能性基因OCT 4和NANOG的启动子去甲基化，从而增加OCT 4和NANOG的表达长达3天，并创建干细胞的治疗窗口期。
结论
总之，nsPEFs可以通过表观遗传调控促进MSCs分化，可能是未来干细胞应用的安全有效策略。
Background
Multiple strategies have been proposed to promote the differentiation potential of mesenchymal stem cells (MSCs), which is the fundamental property in tissue formation and regeneration. However, these strategies are relatively inefficient that limit the application. In this study, we reported a novel and efficient strategy, nanosecond pulsed electric fields (nsPEFs) stimulation, which can enhance the trilineage differentiation potential of MSCs, and further explained the mechanism behind.
Methods
We used histological staining to screen out the nsPEFs parameters that promoted the trilineage differentiation potential of MSCs, and further proved the effect of nsPEFs by detecting the functional genes. In order to explore the corresponding mechanism, we examined the expression of pluripotency genes and the methylation status of their promoters. Finally, we targeted the DNA methyltransferase which was affected by nsPEFs.
Results
The trilineage differentiation of bone marrow-derived MSCs was significantly enhanced in vitro by simply pre-treating with 5 pulses of nsPEFs stimulation (energy levels as 10 ns, 20 kV/cm; 100 ns, 10 kV/cm), due to that the nsPEFs demethylated the promoters of stem cell pluripotency genes OCT4 and NANOG through instantaneous downregulation of DNA methylation transferase 1 (DNMT1), thereby increasing the expression of OCT4 and NANOG for up to 3 days, and created a treatment window period of stem cells.
Conclusions
In summary, nsPEFs can enhance MSCs differentiation via the epigenetic regulation and could be a safe and effective strategy for future stem cell application.