Johkura Kohei, Cui Li, Suzuki Akihiro, Teng Ruifeng, Kamiyoshi Akiko, Okamura Shintaro, Kubota Suguru, Zhao Xu, Asanuma Kazuhiko, Okouchi Yasumitsu, Ogiwara Naoko, Tagawa Yoh ichi, Sasaki Katsunori
Department of Anatomy and Organ Technology, Shinshu University School of Medicine, 3-1-1 Asahi, 390-8621, Matsumoto, Japan.
Cardiovasc Res. 2003 May 1;58(2):435-43. doi: 10.1016/s0008-6363(02)00730-7.
Embryonic stem cell-derived cardiomyocytes are a useful source for cell transplantation into the heart, as well as for tissue engineering of the extracardiac vascular system. The present study was designed to investigate the survival and contractile function of embryonic stem cell-derived cardiomyocytes around large blood vessels to assess the feasibility of their ectopic use for future engineering of cardiovascular tissues.
The mouse embryonic stem cell-derived cardiomyocytes were transplanted into the retroperitoneum of the adult nude mice, and the myocardial tissues that developed were characterized by electrophysiological and histological techniques.
Macroscopic and electrophysiological analyses showed spontaneously contracting transplants in the host retroperitoneum 7 and 30 days after transplantation. Immunohistochemistry detected developing cardiomyocytes in the transplants on Day 7, which formed the myocardial tissues. They were positive for cardiac troponin I, cadherin, connexin 43, and proliferating cell nuclear antigen, but negative for alpha-smooth muscle actin. Vascular formation was discernible in the transplant tissues. By Day 30, more mature myocardial tissues had been established in the transplants. Electron microscopic study emphasized that the transplant tissues comprised cardiomyocytes, in which myofibrils with organized sarcomeres were observed. Desmosomes, fasciae adherens and gap junctions were evident in the cellular junctions.
The cardiomyocytes derived from the mouse ES cells were demonstrated to be viable and function in the ectopic site of the host retroperitoneum up to Day 30, following a process of proliferation and differentiation. Vascularization and host perfusion beneficial for the survival of the cardiomyocytes occurred in the transplants.
胚胎干细胞衍生的心肌细胞是心脏细胞移植以及心外血管系统组织工程的有用细胞来源。本研究旨在调查胚胎干细胞衍生的心肌细胞在大血管周围的存活情况和收缩功能,以评估其异位用于未来心血管组织工程的可行性。
将小鼠胚胎干细胞衍生的心肌细胞移植到成年裸鼠的腹膜后,并用电生理和组织学技术对发育形成的心肌组织进行表征。
宏观和电生理分析显示,移植后7天和30天,宿主腹膜后有自发收缩的移植物。免疫组织化学检测到移植后第7天移植物中有发育中的心肌细胞,这些细胞形成了心肌组织。它们对心肌肌钙蛋白I、钙黏蛋白、连接蛋白43和增殖细胞核抗原呈阳性,但对α-平滑肌肌动蛋白呈阴性。移植组织中可见血管形成。到第30天,移植物中已建立了更成熟的心肌组织。电子显微镜研究强调,移植组织由心肌细胞组成,其中观察到具有有组织肌节的肌原纤维。细胞连接处可见桥粒、黏着带和缝隙连接。
来自小鼠胚胎干细胞的心肌细胞在增殖和分化过程后,在宿主腹膜后的异位部位直至第30天被证明是有活力且能发挥功能的。移植组织中发生了有利于心肌细胞存活的血管化和宿主灌注。