激活素A和骨形成蛋白-4诱导人羊膜上皮细胞转分化为心肌样细胞的实验研究  

Activin A and BMP-4 Induce Cardiomyocyte-like Cells Differentiation of Human Amniotic Epithelial Cells

作　　者：韩晓宇[1,2] 万琪[3] 吴文超[1] 郑艾[3] 李良[1,4] 刘小菁[1,2] 

机构地区：[1]四川大学华西医院心血管疾病研究室,成都610041 [2]四川大学华西医院心血管内科,成都610041 [3]四川大学华西第二医院妇产科,成都610041 [4]四川大学华西医学中心生物医学工程研究室,成都610041

出　　处：《生物医学工程学杂志》2011年第6期1217-1222,共6页Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目(30870598;10972149)

摘　　要：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。Amniotie epithelial cells （AECs） have been expected to be a good cell source for stem cell-based cardiac repair. Activin A signaling is required for cardiac differentiation in human embryonic stem cells （ESCs）, and bone morphogenetic protein-4 （BMP-4） is an important regulator that controls stem cell fates. Previous study has established an efficient protocol to generate cardiomyocytes from human ESCs via induction with Aetivin A and BMP-4. The aim of present study was to test the hypothesis that Activin A and BMP-4 could also induce AECs to differentiate into cardiomyocytes in vitro. Human AECs （hAECs） were isolated from human term placenta by trypsin digestion according to the previous reports. Freshly isolated hAECs were examined to detect the expression of cytokeratin 19 by immunoeytochemistry. High-density undifferentiated hAECs at passage 1 were sequential treated with 100 ng/ ml human recombinant Actvin A and 10ng/ml BMP-4. The expression of cardiac-specific genes was examined before and after in vitro induction of cellular differentiation. Freshly isolated hAEC could express cytokeratin 19, the specific marker of epithelial cells. The data showed that hAECs treated with Aetivin A and BMP-4 were able to express cardiac-specific genes, including Nkx2.5 and a-actinin. Our results demonstrated that Activin A and BMP-4 could induce cardiomyocyte differentiation of hAECs, which might be a novel approach to induce differentiation of AECs into cardiomyocytes-like cells.

关 键 词：羊膜上皮细胞 激活素A 骨形成蛋白-4 心肌样细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]