首页> 外文期刊>International Journal of Clinical and Experimental Pathology >Differentiation induction of mouse cardiac stem cells into sinus node-like cells by co-culturing with sinus node
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Differentiation induction of mouse cardiac stem cells into sinus node-like cells by co-culturing with sinus node

机译:与窦房结共培养诱导小鼠心脏干细胞分化为窦房结样细胞

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Sinus nodal cells can generate a diastolic or “pacemaker” depolarization at the end of an action potential driving the membrane potential slowly up to the threshold for firing the next action potential. It has been proved that adult cardiac stem cells (CSCs) can differentiate into sinus nodal cells by demethylating agent. However, there is no report about adult CSCs-derived sinus nodal cells with pacemaker current (the funny current, I f). In this study, we isolated the mouse adult CSCs from mouse hearts by the method of tissue explants adherence. The expression of c-kit protein indicated the isolation of CSCs. Then we co-cultured mouse CSCs with mouse sinus node tissue to induce the differentiation of these CSCs into sinus node-like cells, which was proved by identifying the enhanced expression of marker proteins cTnI, cTnT and α-Actinin with Immunofluorescence staining. At the same time, with whole-cell patch-clamp we detected the I f current, which can be blocked by CsCl, in these differentiated cells. In conclusion, by confirming specific I f current in the induced node-like cells, our work shows a method inducing differentiation of CSCs into sinus node-like cells, which can provide helpful information for the further research on sick sinus syndrome.
机译:窦房结细胞可在动作电位结束时产生舒张或“起搏器”去极化,从而将膜电位缓慢驱动至触发下一个动作电位的阈值。业已证明,成年心脏干细胞(CSCs)可以通过去甲基化剂分化为窦房结细胞。然而,没有关于起搏器电流(有趣的电流,I f)的成年CSC来源的窦房结细胞的报道。在这项研究中,我们通过组织外植体粘附的方法从小鼠心脏中分离了小鼠成年CSC。 c-kit蛋白的表达表明CSC的分离。然后,我们将小鼠CSCs与小鼠窦房结组织共培养,以诱导这些CSCs向窦房结样细胞的分化,这通过免疫荧光染色鉴定了标记蛋白cTnI,cTnT和α-Actinin的增强表达来证明。同时,通过全细胞膜片钳,我们在这些分化的细胞中检测到了I f电流,该电流可以被CsCl阻断。总之,通过确定诱导的节点样细胞中特定的I f电流,我们的工作表明了一种诱导CSCs分化为窦房结样细胞的方法,这可为进一步研究病态窦房结综合征提供有用的信息。

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