Endothelial colony-forming cells for preparing prevascular three-dimensional cell-dense tissues using cell-sheet engineering

被引:20
作者
Sasagawa, Tadashi [1 ]
Shimizu, Tatsuya [1 ]
Yamato, Masayuki [1 ]
Okano, Teruo [1 ]
机构
[1] Tokyo Womans Med Univ TWIns, Inst Adv Biomed Engn & Sci, Tokyo, Japan
基金
日本学术振兴会;
关键词
endothelial colony-forming cells; cell-sheet; cell-sheet stacking 3D sandwich co-culture; prevascularization; neovascularization; functional anastomosis; UMBILICAL-CORD BLOOD; PROGENITOR CELLS; IN-VIVO; VITRO; VESSELS; ANGIOGENESIS; VASCULATURE; FIBROBLASTS; FABRICATION; CONTRIBUTE;
D O I
10.1002/term.1858
中图分类号
Q813 [细胞工程];
学科分类号
摘要
Vascular-derived endothelial cell (EC) network prefabrication in three-dimensional (3D) tissue constructs before transplantation is useful for inducing functional anastomosis with the host vasculature. However, the clinical application of ECs is limited by cell isolation from the existing vasculature, because of the requirement for invasive biopsies and difficulty in obtaining a sufficient number of cells. Endothelial colony-forming cells (ECFCs), which are a subtype of endothelial progenitor cells in the blood, have a strong proliferative and vasculogenic potential. This study attempted to fabricate prevascular 3D cell-dense tissue constructs using cord blood-derived ECFCs and evaluate the in vivo angiogenic potential of these constructs. Human umbilical vascular endothelial cells (HUVECs) were also used in comparison with ECFCs, which were sandwiched between two human dermal-derived fibroblast (FB) sheets using a fibrin-coated cell-sheet manipulator. The inserted ECFCs in double-layered FB sheets were cultured for 3 days, resulting in the formation of network structures similar to those of HUVECs. Additionally, when ECFCs were sandwiched with three FB sheets, a lumen structure was found in the triple-layered cell-sheet constructs at 3 days after co-culture. These constructs containing ECFCs were transplanted into the subcutaneous tissue of immune-deficient rats. One week after transplantation, ECFC-lined functional microvessels containing rat erythrocytes were observed in the same manner as transplanted HUVEC-positive grafts. These results suggest that ECFCs might become an alternative cell source for fabricating a prevascular structure in 3D cell-dense tissue constructs for clinical application. Copyright (C) 2013 John Wiley & Sons, Ltd.
引用
收藏
页码:739 / 747
页数:9
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