SHIMIZU Tatsuya    Department   Research Institutes and Facilities, Research Institutes and Facilities    Position   Professor
Article types	Original article
Language	English
Peer review	Peer reviewed
Title	Endothelial colony-forming cells for preparing prevascular three-dimensional cell-dense tissues using cell-sheet engineering.
Journal	Formal name：Journal of tissue engineering and regenerative medicine Abbreviation：J Tissue Eng Regen Med ISSN code：(1932-7005)1932-6254(Linking)
Domestic / Foregin	Foregin
Volume, Issue, Page	10(9),pp.739-747
Author and coauthor	Sasagawa Tadashi†, Shimizu Tatsuya, Yamato Masayuki, Okano Teruo*
Authorship	2nd author
Publication date	2016/09
Summary	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 © 2013 John Wiley & Sons, Ltd.
DOI	10.1002/term.1858
PMID	24668945