Effects of angiogenic factors and 3D-microenvironments on vascularization within sandwich cultures

• Published in: Biomaterials Vol. 35; no. 17; pp. 4739 - 4748
• Main Authors: Nishiguchi, Akihiro, Matsusaki, Michiya, Asano, Yoshiya, Shimoda, Hiroshi, Akashi, Mitsuru
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Ltd 01.06.2014
• Subjects: Advanced Basic Science; Angiogenesis; Artificial tissue engineering; blood; Cell Hypoxia; Cytokine; Dentistry; ECM (extracellular matrix); Endothelial cell; endothelial cells; Endothelial Cells - cytology; Endothelial Cells - metabolism; Fibroblast Growth Factor 2 - metabolism; Fibroblasts - cytology; Fibroblasts - metabolism; Hepatocyte Growth Factor - metabolism; Human Umbilical Vein Endothelial Cells; Humans; lymph; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - metabolism; Neovascularization, Physiologic; secretion; stem cells; tissue engineering; Tissue Engineering - methods; Tissue Scaffolds - chemistry; tissues; Vascular Endothelial Growth Factor A - metabolism; ECM (extracellular matrix); Angiogenesis; Endothelial cell; Artificial tissue engineering; Cytokine
• ISSN: 0142-9612; 1878-5905; 1878-5905
• DOI: 10.1016/j.biomaterials.2014.01.079

The in vitro fabrication of vascularized tissue is a key challenge in tissue engineering, but little is known about the mechanisms of blood-capillary formation. Here we investigated the mechanisms of in vitro vascularization using precisely-controlled 3D-microenvironments constructed by a sandwich culture using the cell-accumulation technique. 3D-microenvironments controlled at the single layer level showed that sandwich culture between more than 3 fibroblast-layers induced tubule formation. Moreover, the secretion of angiogenic factors increased upon increasing the number of sandwiching layers, which induced highly dense tubular networks. We found that not only angiogenic factors, but also the 3D-microenvironments of the endothelial cells, especially apical side, played crucial roles in tubule formation in vitro. Based on this knowledge, the introduction of blood and lymph capillaries into mesenchymal stem cell (MSC) tissues was accomplished. These findings would be useful for the in vitro vascularization of various types of engineered organs and studies on angiogenesis.