The long-term differentiation of embryonic stem cells into cardiomyocytes: an indirect co-culture model.
The long-term differentiation of embryonic stem cells into cardiomyocytes: an indirect co-culture model.
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DOI：
10.1371/journal.pone.0055233
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Zheng QS
中科院分区：
文献类型：
--
作者：
Ou DB;Zeng D;Jin Y;Liu XT;Teng JW;Guo WG;Wang HT;Su FF;He Y;Zheng QS
胚胎干细胞（Embryonic Stem cells，ESCs）在体外可分化为心肌细胞（myocardial cells，CM），但分化水平低。在这里，我们描述了一种简单的共培养模型，通过市售的Millicell™悬挂细胞培养插入物来控制ESC向CM的长期分化。将小鼠胚胎干细胞悬滴培养形成胚状体（EBs），用0.1mmol/L抗坏血酸诱导其向CM分化。在间接共培养系统中，通过悬挂细胞培养插入物（具有1 μm孔的PET膜）将EB与表皮角质形成细胞（EK）或新生儿CM（NCM）共培养。评价了在延长的培养过程中ESC衍生的CM的分子表达和功能特性。在ESC分化过程中，NCMs共培养的EB面积收缩率显著高于未共培养的EB和EKs共培养的EB。在NCMs共培养模型中，ESC来源的CM的功能维持更加突出。这些结果表明，NCMs共培养促进ESC分化，并对细胞生长和分化具有进一步的影响。我们认为，在共培养系统中，ESCs向CM的分化效率的提高不是由于共培养对细胞分化的直接影响，而是通过影响细胞增殖和长期功能维持的信号作用。
Embryonic Stem Cells (ESCs) can differentiate into cardiomyocytes (CMs) in vitro but the differentiation level from ESCs is low. Here we describe a simple co-culture model by commercially available Millicell™ hanging cell culture inserts to control the long-term differentiation of ESCs into CMs. Mouse ESCs were cultured in hanging drops to form embryoid bodies (EBs) and treated with 0.1 mmol/L ascorbic acid to induce the differentiation of ESCs into CMs. In the indirect co-culture system, EBs were co-cultured with epidermal keratinocytes (EKs) or neonatal CMs (NCMs) by the hanging cell culture inserts (PET membranes with 1 µm pores). The molecular expressions and functional properties of ESC-derived CMs in prolonged culture course were evaluated. During time course of ESC differentiation, the percentages of EBs with contracting areas in NCMs co-culture were significantly higher than that without co-culture or in EKs co-culture. The functional maintenance of ESC-derived CMs were more prominent in NCMs co-culture model. These results indicate that NCMs co-culture promote ESC differentiation and has a further effect on cell growth and differentiation. We assume that the improvement of the differentiating efficiency of ESCs into CMs in the co-culture system do not result from the effect of co-culture directly on cell differentiation, but rather by signaling effects that influence the cells in proliferation and long-term function maintenance.
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