Takei Takayuki, Sakai Shinji, Yokonuma Toru, Ijima Hiroyuki, Kawakami Koei
Department of Chemical Engineering, Faculty of Engineering, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka 819-0385, Japan.
Biotechnol Prog. 2007 Jan-Feb;23(1):182-6. doi: 10.1021/bp060152j.
One possible strategy for creating three-dimensional (3D) tissue-engineered organs in vitro is to develop a vasculature for sufficient transport of oxygen and nutrients within these constructs. Here, we describe a novel technique to fabricate endothelialized tubes with predetermined 3D configuration, as a starting point for self-developing capillary-like networks in vitro. Calcium-alginate hydrogel fibers of ca. 250 and 500 mum in diameter, enclosing bovine carotid artery vascular endothelial cells (BECs), were used as templates for endothelialized tubes. Fibers were prepared by extruding a 2% (w/v) sodium alginate solution containing BECs into a 100 mM calcium chloride solution flowing in the same direction. Fibers were embedded in type I collagen gels and enzymatically degraded by alginate lyase, resulting in channels with predetermined 3D configuration filled with a BEC suspension. Cells attached to and covered the surfaces of the channels. Exposing the cells to medium containing basic fibroblast growth factor resulted in their migration into the ambient collagen gel and self-assembly into capillary-like structures. These results demonstrate that using artificial endothelialized tubes with predetermined 3D configuration, as a starting point for a self-developing capillary-like network, could be potentially useful for constructing 3D tissue-engineered organs.
在体外创建三维（3D）组织工程器官的一种可能策略是构建脉管系统，以便在这些构建体中充分运输氧气和营养物质。在此，我们描述了一种制造具有预定3D构型的内皮化管的新技术，作为体外自发生成类毛细血管网络的起点。直径约250和500微米的海藻酸钙水凝胶纤维包裹着牛颈动脉血管内皮细胞（BECs），用作内皮化管的模板。通过将含有BECs的2%（w/v）海藻酸钠溶液挤出到同向流动的100 mM氯化钙溶液中来制备纤维。将纤维包埋在I型胶原凝胶中，并用海藻酸裂解酶进行酶解，从而形成具有预定3D构型的通道，通道中充满了BEC悬浮液。细胞附着并覆盖在通道表面。将细胞暴露于含有碱性成纤维细胞生长因子的培养基中，导致它们迁移到周围的胶原凝胶中并自组装成类毛细血管结构。这些结果表明，使用具有预定3D构型的人工内皮化管作为自发生成类毛细血管网络的起点，可能对构建3D组织工程器官有潜在用途。
