Miskon Azizi, Ehashi Tomo, Mahara Atsushi, Uyama Hiroshi, Yamaoka Tetsuji
Department of Biomedical Engineering, Advanced Medical Engineering Center, National Cardiovascular Center Research Institute, Suita, Osaka, Japan.
J Artif Organs. 2009;12(2):111-7. doi: 10.1007/s10047-009-0449-4. Epub 2009 Jun 18.
Stem cell-based therapy in cardiac tissue engineering is an emerging field that shows great potential for treating heart diseases. However, even preliminary issues, such as the ideal niche for cardiomyocytes, have not been clarified yet. In the present study, the effects of extracellular matrix (ECM) components on the beating duration of neonatal rat cardiomyocytes (RCMs) and on the cardiac differentiation of P19.CL6 carcinoma stem cells were studied. RCMs were cultured on gelatin-, fibronectin-, and collagen type I-coated dishes and on noncoated polystyrene dishes, and their beating rate, beating duration, and cardiac gene expression were evaluated. The beating period and the expression of troponin T type-2 (TNNT2) and troponin C type-1 (TNNC1) of cardiomyocytes cultured on gelatin-coated dishes were longer and higher than for those on dishes with other coatings. For the cardiac differentiation of P19.CL6 cells, troponin T type-2 expression on gelatin- and fibronectin-coated dishes was five times that on collagen type I-coated dishes or polystyrene dishes 11 days after induction. These results indicate that a gelatin-coated surface has a high ability not only to maintain the cardiac phenotype but also to enhance cardiac differentiation.
基于干细胞的心脏组织工程疗法是一个新兴领域，在治疗心脏病方面显示出巨大潜力。然而，即使是一些初步问题，如心肌细胞的理想微环境，尚未得到阐明。在本研究中，研究了细胞外基质（ECM）成分对新生大鼠心肌细胞（RCMs）搏动持续时间以及对P19.CL6癌干细胞心脏分化的影响。将RCMs培养在明胶、纤连蛋白和I型胶原包被的培养皿以及未包被的聚苯乙烯培养皿上，并评估其搏动率、搏动持续时间和心脏基因表达。在明胶包被培养皿上培养的心肌细胞的搏动周期以及肌钙蛋白T 2型（TNNT2）和肌钙蛋白C 1型（TNNC1）的表达，比在其他包被培养皿上培养的心肌细胞更长、更高。对于P19.CL6细胞的心脏分化，诱导11天后，明胶和纤连蛋白包被培养皿上的肌钙蛋白T 2型表达是I型胶原包被培养皿或聚苯乙烯培养皿上的五倍。这些结果表明，明胶包被表面不仅具有维持心脏表型的高能力，而且具有增强心脏分化能力。
