title: Expression profiles of angiogenesis-related proteins in prevascular three-dimensional tissues using cell-sheet engineering.
abstract: The prefabrication of endothelial cell network assembly (ECNA) in tissue-engineer multi-layered cell-sheets, known as in vitro prevascularization, is beneficial strategy for inducing anastomosis with the host vasculature after transplantation. However, the mechanisms of neovascularization via transplanted prevascular cell-sheets are unknown. This study investigated neovascularization process and angiogenesis-related protein secretion by prevascular cell-sheets. Prevascular (ECNA-positive) double-layered fibroblast (FB) sheets were created by sandwiching human aortic endothelial cells (HAECs) between two human dermal FB sheets. As the ECNA-negative control, FBs-sandwiching double-layered FB sheets were used. Two types of cell-sheets were subcutaneously transplanted into immune-deficient rats. At 3 days after transplantation, induction of the newly-formed microvessels near the host vasculature was observed in the ECNA-positive cell-sheet. In contrast, no neovessel was observed in the ECNA-negative cell-sheet at 1 week after transplantation. Consequently, the secretion of angiogenesis-related proteins in conditioned media of each cell-sheet cultured for 3 days were compared. The levels of hepatocyte growth factor (HGF), placenta growth factor (PlGF) and matrix metalloproteinase-9 (MMP-9) significantly increased in the ECNA-positive cell-sheets. These results suggested that these molecules might involve in neovascularization after the transplantation of prevascular cell-sheets. These findings may contribute to understanding its mechanism.
authors.affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo Woman's Medical University (TWIns), Tokyo, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Sasagawa
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Shimizu
authors.forename: Masayuki
authors.initials: M
authors.lastname: Yamato
authors.forename: Teruo
authors.initials: T
authors.lastname: Okano
mesh_terms: D002478:Cells, Cultured; D018920:Coculture Techniques; D017077:Culture Media, Conditioned; D020869:Gene Expression Profiling; D006801:Humans; D018919:Neovascularization, Physiologic; D011506:Proteins / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D023822:Tissue Engineering; D054457:Tissue Scaffolds
chemical_list: D017077:Culture Media, Conditioned; D011506:Proteins
keywords: Angiogenesis-related protein; Cell-sheet; Endothelial cell; Endothelial cell network assembly; Neovascularization; Sandwich co-culture