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近年来羊膜上皮细胞(AECs)被认为是心肌细胞移植较好的种子细胞之一。研究表明,激活素A(ActivinA)和骨形成蛋白-4(BMP-4)能够诱导人胚胎干细胞向心肌细胞分化。本研究通过体外分离、培养并鉴定人AEC后,采用Activin A和BMP-4作为诱导剂,观察它们能否诱导AEC向心肌样细胞转分化。应用定量RT-PCR、Western blot、免疫细胞化学等方法来检测基因及蛋白表达变化。结果显示,人AEC能够表达上皮细胞特异性蛋白角蛋白19(CK19)。经Activin A和BMP-4共同诱导培养14d后,AEC心肌特异性转录因子Nkx2.5和特异性结构蛋白α-actinin的mRNA表达水平明显升高,同时α-actinin的蛋白表达水平亦明显增加。本实验提示,ActivinA和BMP-4在体外能够诱导人AEC向心肌样细胞转分化,同时该诱导方法可能成为诱导AEC向心肌样细胞转分化的一个新方法。
In recent years, amniotic membrane epithelial cells (AECs) are considered as one of the better seed cells for cardiomyocyte transplantation. Studies have shown that activin A and BMP-4 can induce human embryonic stem cells to differentiate into cardiomyocytes. In this study, we isolated and cultured human AECs and identified them as activators of activin A and BMP-4 as inducers to observe whether they can induce AEC transdifferentiation into cardiomyocyte-like cells. Quantitative RT-PCR, Western blot, immunocytochemistry and other methods to detect changes in gene and protein expression. The results showed that human AEC can express epithelial cell-specific protein keratin 19 (CK19). After induced by Activin A and BMP-4 for 14 days, the expression of myocardial specific transcription factor Nkx2.5 and specific structural protein α-actinin in AEC increased significantly, and the protein expression of α-actinin also increased significantly. This experiment suggests that ActivinA and BMP-4 can induce human AEC transdifferentiation into cardiomyocyte-like cells in vitro, and this induction method may become a new method to induce transdifferentiation of AEC into cardiomyocyte-like cells.