title: Generation of functional endothelial-like cells from adult mouse germline-derived pluripotent stem cells.
abstract: Functional endothelial cells and their progenitors are required for vascular development, adequate vascular function, vascular repair and for cell-based therapies of ischemic diseases. Currently, cell therapy is limited by the low abundance of patient-derived cells and by the functional impairment of autologous endothelial progenitor cells (EPCs). In the present study, murine germline-derived pluripotent stem (gPS) cells were evaluated as a potential source for functional endothelial-like cells. Cells displaying an endothelial cell-like morphology were obtained from gPS cell-derived embryoid bodies using a combination of fluorescence-activated cell sorting (FACS)-based selection of CD31-positive cells and their subsequent cultivation on OP9 stromal cells in the presence of VEGF-A. Real-time reverse transcriptase polymerase chain reaction, FACS analysis and immunofluorescence staining showed that the gPS cell-derived endothelial-like cells (gPS-ECs) expressed endothelial cell-specific markers including von Willebrand Factor, Tie2, VEGFR2/Flk1, intercellular adhesion molecule 2 and vascular endothelial-cadherin. The high expression of ephrin B2, as compared to Eph B4 and VEGFR3, suggests an arterial rather than a venous or lymphatic differentiation. Their capability to take up Dil-conjugated acetylated low-density lipoprotein and to form capillary-like networks on matrigel confirmed their functionality. We conclude that gPS cells could be a novel source of endothelial cells potentially suitable for regenerative cell-based therapies for ischemic diseases.
authors: Department of Cell and Developmental Biology, Max Planck Institute of Molecular Biomedicine, Röntgenstr. 20, D-48149 Münster, Germany.
authors: Julee
authors: J
authors: Kim
authors: Department of Cardiovascular Medicine, University Hospital of Münster, Albert-Schweitzer-Campus 1, Bldg. A1, D-48149 Münster, Germany.
authors: Sarah
authors: S
authors: Eligehausen
authors: Department of Cell and Developmental Biology, Max Planck Institute of Molecular Biomedicine, Röntgenstr. 20, D-48149 Münster, Germany.
authors: Martin
authors: M
authors: Stehling
authors: Department of Cardiovascular Medicine, University Hospital of Münster, Albert-Schweitzer-Campus 1, Bldg. A1, D-48149 Münster, Germany.
authors: Sigrid
authors: S
authors: Nikol
authors: Department of Cell and Developmental Biology, Max Planck Institute of Molecular Biomedicine, Röntgenstr. 20, D-48149 Münster, Germany.
authors: Kinarm
authors: K
authors: Ko
authors: Department of Cardiovascular Medicine, University Hospital of Münster, Albert-Schweitzer-Campus 1, Bldg. A1, D-48149 Münster, Germany. Electronic address: johannes.waltenberger@ukmuenster.de.
authors: Johannes
authors: J
authors: Waltenberger
authors: Department of Cardiovascular Medicine, University Hospital of Münster, Albert-Schweitzer-Campus 1, Bldg. A1, D-48149 Münster, Germany. Electronic address: rainer.klocke@ukmuenster.de.
authors: Rainer
authors: R
authors: Klocke
mesh_terms: D053687:Adult Stem Cells / Q000166:cytology* / Q000502:physiology*; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival / Q000502:physiology; D042783:Endothelial Cells / Q000166:cytology* / Q000502:physiology*; D051379:Mice; D023822:Tissue Engineering / Q000379:methods*
keywords: Angiogenesis; Arteriogenesis; Cell-based therapy; Endothelial cells; Germline-derived pluripotent stem cells; Ischemia