Assessment of the ultrastructural and proliferative properties of human embryonic stem cell-derived cardiomyocytes
Assessment of the ultrastructural and proliferative properties of human embryonic stem cell-derived cardiomyocytes
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DOI：
10.1152/ajpheart.00020.2003
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发表时间：
2003-12-01
影响因子：
4.8
通讯作者：
Gepstein, L
中科院分区：
文献类型：
--
作者：
Snir, M;Kehat, I;Gepstein, L
由于缺乏合适的体外模型，对人类心脏组织早期超微结构发育和细胞周期调节的评估受到严重阻碍。在这里，我们描述了可能利用人类胚胎干细胞（ES）系来研究这些过程。利用胚状体（EB）分化系统，分离不同发育阶段的人ES细胞源性心肌细胞，并对其组织形态学、超微结构和增殖特性进行表征。组织形态学分析显示，与早期（10-21天）和中期（21-35天）相比，晚期EBs（>35天）的细胞长度、面积和长宽比增加。同时伴有从不规则的肌原纤维分布到有组织的肌纤维结构的渐进的超微结构发展。心肌细胞增殖，通过心脏特异性抗体和[H-3]胸苷结合或Ki-67免疫标记的双重标记来评估，表明细胞周期逐渐退出。因此，早期心肌细胞（[H-3]胸苷：60+/-10%，Ki-67: 54+/-23%）中阳性染色核的百分比在中期EBs和中期EBs分别下降到36+/-7%和9+/-16%
Assessment of early ultrastructural development and cell-cycle regulation in human cardiac tissue is significantly hampered by the lack of a suitable in vitro model. Here we describe the possible utilization of human embryonic stem cell (ES) lines for investigation of these processes. With the use of the embryoid body (EB) differentiation system, human ES cell-derived cardiomyocytes at different developmental stages were isolated and their histomorphometric, ultrastructural, and proliferative properties were characterized. Histomorphometric analysis revealed an increase in cell length, area, and length-to-width ratio in late-stage EBs (>35 days) compared with early (10-21 days) and intermediate (21-35 days) stages. This was coupled with a progressive ultrastructural development from an irregular myofibrillar distribution to an organized sarcomeric pattern. Cardiomyocyte proliferation, assessed by double labeling with cardiac-specific antibodies and either [H-3] thymidine incorporation or Ki-67 immunolabeling, demonstrated a gradual withdrawal from cell cycle. Hence, the percentage of positively stained nuclei in early-stage cardiomyocytes ([H-3] thymidine: 60+/-10%, Ki-67: 54+/-23%) decreased to 36+/-7% and 9+/-16% in intermediate-stage EBs and to