Cardiac differentiation of P19CL6 cells by oxytocin
Cardiac differentiation of P19CL6 cells by oxytocin
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DOI：
10.1016/j.ijcard.2008.01.046
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发表时间：
2009-05-01
影响因子：
3.5
通讯作者：
Mowla, Seyed Javad
中科院分区：
文献类型：
--
作者：
Fathi, Fardin;Murasawa, Satoshi;Mowla, Seyed Javad
背景：已有研究表明，P19胚胎癌（EC）细胞在催产素（OT）的作用下可分化为搏动的心肌细胞。已经表明二甲基亚砜（DMSO）通过催产素/催产素受体途径起作用，因为催产素受体拮抗剂不仅阻断催产素诱导的心肌细胞分化，而且阻断DMSO诱导的分化。本研究以P19 CL 6细胞为实验材料，观察OT对P19 CL 6细胞的分化能力。然后将OT作为诱导剂添加到培养基中。用1%DMSO处理的细胞用作阳性对照组。分化的细胞进行了形态学和免疫细胞化学，以及通过RT-PCR评价。结果：P19 CL 6细胞聚集成团后可分化为心肌细胞，而单层细胞不能分化并表达特异性心肌标志基因。在对照组中，无论是聚集体细胞还是单层细胞都可以被DMSO诱导分化为心肌细胞。此外，表达GFP的P19 CL 6细胞在用OT处理时有效地分化为搏动的心肌细胞。所有评估的结果证实，分化的细胞是心肌cardiomyocyte.Conclusions：我们得出结论，胚状体的形成（细胞聚集）是必要的P19 CL 6细胞分化为心肌细胞时，使用OT作为诱导剂。此外，由于高分化效率，来源于P19 CL 6细胞的表达GFP的心肌细胞具有用于实验模型中的再生疗法的潜力。(C)2008爱思唯尔爱尔兰有限公司保留所有权利。
Background: It has been reported that P19 embryonal carcinoma (EC) cells differentiate into beating cardiomyocytes under the action of oxytocin (OT). It has been suggested that dimethylsulfoxide (DMSO) acts via the oxytocin/oxytocin receptor pathway because an oxytocin receptor antagonist not only blocks oxytocin-induced cardiomyocyte differentiation, but also blocks DMSO-induced differentiation. In this study, the differentiation ability of OT was tested using P19CL6 cells.Methods: P19CL6 cells were cultured as a confluent monolayer and aggregated cells. OT was then added to culture media as an inducing agent. The cells treated with 1% DMSO were used as a positive control group. Differentiated cells were evaluated morphologically and immunocytochemically, as well as by RT-PCR. In addition, a stable line of green fluorescent protein (GFP)-expressing P19CL6 cells were differentiated into beating cardiomyocytes by OT.Results: Aggregated P19CL6 cells could be differentiated into cardiomyocytes, whereas monolayer cells could not differentiate and express specific cardiac muscle marker genes. In the control group, both aggregates and monolayer cells could be differentiated into cardiomyocytes by DMSO. In addition, GFP-expressing P19CL6 cells differentiated efficiently into beating cardiomyocytes when treated with OT. The results of all evaluations confirmed that the differentiated cells were cardiomyocytes.Conclusions: We concluded that embryoid body formation (cell aggregation) is necessary for the differentiation of P19CL6 cells into cardiomyocytes when using OT as an inducer agent. Furthermore, because of the high rate of differentiation efficiency, GFP-expressing cardiomyocytes derived from P19CL6 cells have the potential to be used for regenerative therapies in experimental models. (C) 2008 Elsevier Ireland Ltd. All rights reserved.