title: Localized Gene Expression Analysis during Sprouting Angiogenesis in Mouse Embryoid Bodies Using a Double Barrel Carbon Probe.
abstract: The mouse embryonic stem (ES) cell-derived angiogenesis model is widely used as a 3D model, reproducing cell-cell interactions in the living body. Previously, many methods to analyze localized cellular function, including in situ hybridization and laser capture microdissection, have been reported. In this study, we achieved a collection of localized cells from the angiogenesis model in hydrogel. The gene expression profiles of the endothelial cells derived from mouse ES cells were evaluated. First, we collected localized cells from the live tissue model embedded in hydrogel using the double barrel carbon probe (DBCP) and quantified mRNA expression. Second, we found that vascular marker genes were expressed at a much higher level in sprouting vessels than in the central core of the embryoid body because the cells in sprouting vessels might significantly differentiate into endothelial linages, including tip/stalk cells. Third, the gene expression levels tended to be different between the top and middle regions in the sprouting vessel due to the difference in the degree of differentiation in these regions. At the top region of the vessel, both the tip and stalk cells were present. The cells in the middle region became more mature. Collectively, these results show that DBCP is very useful for analyzing localized gene expression in cells collected from 3D live tissues embedded in hydrogel. This technique can be applied to comprehensive gene expression analyses in the medical field.
authors.affiliation: Graduate School of Environmental Studies, Tohoku University , Sendai 980-8579, Japan.
authors.forename: Hidenori
authors.initials: H
authors.lastname: Ito
authors.affiliation: Graduate School of Environmental Studies, Tohoku University , Sendai 980-8579, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Nashimoto
authors.affiliation: WPI-Advanced Institute for Materials Research, Tohoku University , Sendai 980-8577, Japan.
authors.forename: Yuanshu
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Graduate School of Environmental Studies, Tohoku University , Sendai 980-8579, Japan.|WPI-Advanced Institute for Materials Research, Tohoku University , Sendai 980-8577, Japan.|PRESTO, JST, Saitama 332-0012, Japan.
authors.forename: Yasufumi
authors.initials: Y
authors.lastname: Takahashi
authors.affiliation: Graduate School of Environmental Studies, Tohoku University , Sendai 980-8579, Japan.
authors.forename: Kosuke
authors.initials: K
authors.lastname: Ino
authors.affiliation: Graduate School of Environmental Studies, Tohoku University , Sendai 980-8579, Japan.
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Shiku
authors.affiliation: Graduate School of Environmental Studies, Tohoku University , Sendai 980-8579, Japan.|WPI-Advanced Institute for Materials Research, Tohoku University , Sendai 980-8577, Japan.
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Matsue
mesh_terms: D000818:Animals; D002244:Carbon / Q000737:chemistry / Q000378:metabolism*; D002454:Cell Differentiation; D002478:Cells, Cultured; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism*; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods*; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D023421:Models, Animal; D018919:Neovascularization, Physiologic / Q000235:genetics*
chemical_list: D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D002244:Carbon