Shox2 influences mesenchymal stem cell fate in a co-culture model in vitro.
Shox2 influences mesenchymal stem cell fate in a co-culture model in vitro.
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DOI：
10.3892/mmr.2016.5306
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发表时间：
2016-07
影响因子：
3.4
通讯作者：
Song Z
中科院分区：
文献类型：
--
作者：
Feng Y;Yang P;Luo S;Zhang Z;Li H;Zhu P;Song Z
窦房结功能障碍是一种常见的心血管疾病，细胞源性生物起搏器的研制一直是心脏电生理研究的热点。生物起搏器治疗的目的是产生SAN样细胞，其表现出SAN的自发活动特征。Shox 2是一种早期心脏转录因子，在窦房结（SAN）的形成和分化中起关键作用。本研究旨在通过在犬间充质干细胞（cMSCs）中过表达Shox 2以诱导类似于天然起搏细胞的表型来改善起搏功能。为了实现这一目的，用慢病毒pLentis-mShox 2-red荧光蛋白转染cMSCs，然后与大鼠新生心肌细胞（RNCM）在体外共培养5-7天。研究Shox 2过表达调控cMSCs向起搏样细胞分化的可行性。逆转录-定量聚合酶链反应和蛋白质印迹显示，Shox 2转染的cMSCs表达高水平的T盒3，超极化激活的环核苷酸门控阳离子通道和连接蛋白45基因，参与SAN的发展，和低水平的工作心肌基因，Nkx2.5和连接蛋白43。此外，Shox 2转染的cMSC能够以比对照细胞更快的速率起搏RNCM。总之，这些数据表明，在体外共培养模型中，在cMSC中过表达Shox 2可以大大增强起搏表型。
Sinoatrial node (SAN) dysfunction is a common cardiovascular problem, and the development of a cell sourced biological pacemaker has been the focus of cardiac electrophysiology research. The aim of biological pacemaker therapy is to produce SAN-like cells, which exhibit spontaneous activity characteristic of the SAN. Short stature homeobox 2 (Shox2) is an early cardiac transcription factor and is crucial in the formation and differentiation of the sinoatrial node (SAN). The present study aimed to improve pacemaker function by overexpression of Shox2 in canine mesenchymal stem cells (cMSCs) to induce a phenotype similar to native pacemaker cells. To achieve this objective, the cMSCs were transfected with lentiviral pLentis-mShox2-red fluorescent protein, and then co-cultured with rat neonatal cardiomyocytes (RNCMs) in vitro for 5–7 days. The feasibility of regulating the differentiation of cMSCs into pacemaker-like cells by Shox2 overexpression was investigated. Reverse transcription-quantitative polymerase chain reaction and western blotting showed that Shox2-transfected cMSCs expressed high levels of T box 3, hyperpolarization-activated cyclic nucleotide-gated cation channel and Connexin 45 genes, which participate in SAN development, and low levels of working myocardium genes, Nkx2.5 and Connexin 43. In addition, Shox2-transfected cMSCs were able to pace RNCMs with a rate faster than the control cells. In conclusion, these data indicate that overexpression of Shox2 in cMSCs can greatly enhance the pacemaker phenotype in a co-culture model in vitro.