title: Tracing behavior of endothelial cells promotes vascular network formation.
abstract: The in vitro formation of network structures derived from endothelial cells in grafts before transplantation contributes to earlier engraftment. In a previous study, endothelial cells migrated to form a net-shaped structure in co-culture. However, the specific network formation behavior of endothelial cells during migration remains unclear. In this study, we demonstrated the tracing behavior and cell cycle of endothelial cells using Fucci-labeled (Fluorescent Ubiquitination-based Cell Cycle Indicator) endothelial cells. Here, we observed the co-culture of Fucci-labeled human umbilical vein endothelial cells (HUVECs) together with normal human dermal fibroblasts (NHDFs) using time-lapse imaging and analyzed by multicellular concurrent tracking. In the G0/G1 period, HUVECs migrate faster than in the S/G2/M period, because G0/G1 is the mobile phase and S/G2/M is the proliferation phase in the cell cycle. When HUVECs are co-cultured, they tend to move randomly until they find existing tracks that they then follow to form clusters. Extracellular matrix (ECM) staining showed that collagen IV, laminin and thrombospondin deposited in accordance with endothelial cell networks. Therefore the HUVECs may migrate on the secreted ECM and exhibit tracing behavior, where the HUVECs migrate toward each other. These results suggested that ECM and a cell phase contributed to form a network by accelerating cell migration.
authors.affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), Tokyo, Japan.
authors.forename: Noriko
authors.initials: N
authors.lastname: Yasuda
authors.affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), Tokyo, Japan.
authors.forename: Hidekazu
authors.initials: H
authors.lastname: Sekine
authors.affiliation: Advanced Business Center, Dai Nippon Printing Co., Ltd., Tokyo, Japan.
authors.forename: Ryoma
authors.initials: R
authors.lastname: Bise
authors.affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), Tokyo, Japan.
authors.forename: Teruo
authors.initials: T
authors.lastname: Okano
authors.affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), Tokyo, Japan. Electronic address: shimizu.tatsuya@twmu.ac.jp.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Shimizu
mesh_terms: D002450:Cell Communication; D059447:Cell Cycle Checkpoints; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D018920:Coculture Techniques; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000502:physiology; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008856:Microscopy, Fluorescence; D018919:Neovascularization, Physiologic; D013997:Time Factors; D059008:Time-Lapse Imaging; D014162:Transfection
chemical_list: D008164:Luminescent Proteins
keywords: Cell cycle; Endothelial cells; Migrate; Vascular network