Human embryonic stem cells can differentiate into myocytes with structural and functional properties of cardiomyocytes
Human embryonic stem cells can differentiate into myocytes with structural and functional properties of cardiomyocytes
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DOI：
10.1172/jci200112131
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发表时间：
2001-08-01
影响因子：
15.9
通讯作者：
Gepstein, L
中科院分区：
文献类型：
--
作者：
Kehat, I;Kenyagin-Karsenti, D;Gepstein, L
由于缺乏合适的体外模型，人类心脏组织发育的研究受到阻碍。我们描述了来自人胚胎干细胞（ES）的心肌细胞的表型特性。将人ES细胞悬浮培养并铺板以形成称为胚状体（EB）的聚集体。8.1%的EB出现自发性收缩区。EB内自发收缩区域的细胞用抗心肌肌球蛋白重链、抗α-辅肌动蛋白、抗结蛋白、抗心肌肌钙蛋白I（抗cTnI）和抗ANP抗体染色呈阳性。电子显微镜显示不同程度的肌原纤维组织，与早期心肌细胞一致。RT-PCR研究证实了几个心脏特异性基因和转录因子的表达。细胞外电描记图的特征在于持续30 +/- 25毫秒的急剧分量，随后是347 +/- 120毫秒的缓慢分量。细胞内Ca 2+瞬变显示持续130 +/- 27毫秒的急剧上升和持续200-300毫秒的弛豫分量。应用异丙肾上腺素和氨甲酰胆碱分别诱导正性和负性变时作用。总之，人ES细胞衍生的心肌细胞显示出早期心肌细胞的结构和功能特性。这一独特的分化系统的建立可能对人类心脏早期分化、功能基因组学、药理学检测、细胞治疗和组织工程的研究产生重要影响。
The study of human cardiac tissue development is hampered by the lack of a suitable in vitro model. We describe the phenotypic properties of cardiomyocytes derived from human embryonic stem (ES) cells. Human ES cells were cultivated in suspension and plated to form aggregates termed embryoid bodies (EBs). Spontaneously contracting areas appeared in 8.1% of the EBs. Cells from the spontaneously contracting areas within EBs were stained positively with anti-cardiac myosin heavy chain, anti-alpha -actinin, anti-desmin, anti-cardiac troponin I (anti-cTnI), and anti-ANP antibodies. Electron microscopy revealed varying degrees of myofibrillar organization, consistent with early-stage cardiomyocytes. RT-PCR studies demonstrated the expression of several cardiac-specific genes and transcription factors. Extracellular electrograms were characterized by a sharp component lasting 30 +/- 25 milliseconds, followed by a slow component of 347 +/- 120 milliseconds. Intracellular Ca2+ transients displayed a sharp rise lasting 130 +/- 27 milliseconds and a relaxation component lasting 200-300 milliseconds. Positive and negative chronotropic effects were induced by application of isoproterenol and carbamylcholine, respectively. In conclusion, the human ES cell-derived cardiomyocytes displayed structural and functional properties of early-stage cardiomyocytes. Establishment of this unique differentiation system may have significant impact on the study of early human cardiac differentiation, functional genomics, pharmacological testing, cell therapy, and tissue engineering.