Polysurgery of cell sheet grafts overcomes diffusion limits to produce thick, vascularized myocardial tissues
Polysurgery of cell sheet grafts overcomes diffusion limits to produce thick, vascularized myocardial tissues
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DOI：
10.1096/fj.05-4715fje
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发表时间：
2006-01-01
期刊：
影响因子：
4.8
通讯作者：
Okano, T
中科院分区：
文献类型：
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作者：
Shimizu, T;Sekine, H;Okano, T
近年来，组织工程领域发展迅速，但血管化不良仍然是生物工程细胞致密组织的主要障碍，由于缺氧、营养不足和废物堆积，限制了构建物的可行尺寸。因此，需要新的技术来制造具有良好组织的血管系统的功能性组织。在本研究中，新生大鼠心肌细胞作为完整的薄片从温度反应培养皿中收获，并堆叠成细胞密集的心肌组织。然而，皮下组织层状细胞片的厚度限制类似于80 μ m（3层）。为了克服这一局限性，我们每隔1、2或3天进行三层移植物的重复移植。两个覆盖的移植物完全同步，在间隔1或2天的病例中，整个组织存活而无坏死。多步骤移植也产生了类似1毫米厚的心肌，具有组织良好的微血管网络。此外，在手术可连接的动脉和静脉上制备的功能性多层移植物显示，通过血管进行完全的移植物灌注，并且通过血管直接吻合成功地进行了移植物的异位移植。这些培养的细胞片整合方法克服了产生厚的血管化组织的长期障碍，为临床修复包括受损心肌在内的各种受损器官提供了一种可能的解决方案。
Recently, the field of tissue engineering has progressed rapidly, but poor vascularization remains a major obstacle in bioengineering cell-dense tissues, limiting the viable size of constructs due to hypoxia, nutrient insufficiency, and waste accumulation. Therefore, new technologies for fabricating functional tissues with a well-organized vasculature are required. In the present study, neonatal rat cardiomyocytes were harvested as intact sheets from temperature-responsive culture dishes and stacked into cell-dense myocardial tissues. However, the thickness limit for layered cell sheets in subcutaneous tissue was similar to 80 mu m (3 layers). To overcome this limitation, repeated transplantation of triple-layer grafts was performed at 1, 2, or 3 day intervals. The two overlaid grafts completely synchronized and the whole tissues survived without necrosis in the 1 or 2 day interval cases. Multistep transplantation also created similar to 1 mm thick myocardium with a well-organized microvascular network. Furthermore, functional multilayer grafts fabricated over a surgically connectable artery and vein revealed complete graft perfusion via the vessels and ectopic transplantation of the grafts was successfully performed using direct vessel anastomoses. These cultured cell sheet integration methods overcome long-standing barriers to producing thick, vascularized tissues, revealing a possible solution for the clinical repair of various damaged organs, including the impaired myocardium.