Highly specialized Epithelial cells
that line the Heart
; Blood vessels
; and lymph vessels, forming the Endothelium
. They are polygonal in shape and joined together by Tight junctions
. The tight junctions allow for variable permeability to specific macromolecules that are transported across the endothelial layer.
1Department of Cellular and Molecular Medicine, Laboratory of Molecular and Cellular Signaling, KU Leuven
Intercellular Ca2+-waves are driven by gap junction channels and hemichannels. Here, we describe a method to measure intercellular Ca2+-waves in cell monolayers in response to a local single-cell mechanical stimulus and its application to investigate the properties and regulation of gap junction channels and hemichannels.
Published July 16, 2013. Keywords: Cellular Biology, Molecular Biology, Medicine, Biomedical Engineering, Biophysics, Immunology, Ophthalmology, Gap Junctions, Connexins, Connexin 43, Calcium Signaling, Ca2+, Cell Communication, Paracrine Communication, Intercellular communication, calcium wave propagation, gap junctions, hemichannels, endothelial cells, cell signaling, cell, isolation, cell culture
1Department of Pulmonary Diseases, Institute for Cardiovascular Research, VU University Medical Center, 2Department of Physiology, Institute for Cardiovascular Research, VU University Medical Center
This protocol reviews Electric Cell-substrate Impedance Sensing, a method to record and analyze the impedance spectrum of adherent cells for the quantification of cell attachment, proliferation, motility, and cellular responses to pharmacological and toxic stimuli. Detection of endothelial permeability and assessment of cell-cell and cell-substrate contacts are emphasized.
Published March 28, 2014. Keywords: Bioengineering, ECIS, Impedance Spectroscopy, Resistance, TEER, Endothelial Barrier, Cell Adhesions, Focal Adhesions, Proliferation, Migration, Motility, Wound Healing
1Klinik und Poliklinik für Anästhesiologie, University of Wurzburg
This method describes how to isolate and immortalize microvascular endothelial cells from mouse brain. We describe a step-by-step protocol starting from the homogenization of brain tissue, digestion steps, seeding and immortalization of the cells. Usually, it takes about five weeks to obtain a homogenous, immortalized microvascular endothelial cell line.
Published August 29, 2012. Keywords: Immunology, Neuroscience, Blood-brain barrier, in vitro cell culture models, brain, microvascular endothelial cells, immortalization, cEND
1Department of Bioengineering, University of Pittsburgh, 2Department of Chemical Engineering, University of Pittsburgh
The current study describes a directed differentiation approach in inducing pancreatic differentiation of human embryonic stem cells. Of great significance is the finding that endothelial cell co-culture mediates maturation of human embryonic stem cell derived pancreatic progenitors into insulin expressing cells.
Published March 27, 2012. Keywords: Stem Cell Biology, Human embryonic stem cells, Endothelial cells, Pancreatic differentiation, Co-culture
JoVE Clinical and Translational Medicine
1Lombardi Comprehensive Cancer Center, Georgetown University
This article describes an in vitro technique for monitoring cancer cells invading through a monolayer of endothelial cells. The data is acquired in real-time as a function of changes in impedance on the surface of electrodes at the well bottom.
Published April 1, 2011. Keywords: Cellular Biology, Invasion, HUVEC, xCELLigence, impedance, real-time, cell-index
1Department of Biomedical Engineering, Duke University, 2School of Medicine, Duke University, 3Department of Surgery, Duke University Medical Center, 4School of Medicine, University of Pennsylvania
A method for seeding titanium blood-contacting biomaterials with autologous cells and testing biocompatibility is described. This method uses endothelial progenitor cells and titanium tubes, seeded within minutes of surgical implantation into porcine venae cavae. This technique is adaptable to many other implantable biomedical devices.
Published September 9, 2011. Keywords: Bioengineering, Stent, Titanium, Thrombosis, Endothelial Progenitor Cell, Endothelium, Biomaterial, Biocompatibility, Bioengineering, Translational Medicine, Vascular Surgery, Porcine
1Division of Cardiovascular Medicine, Stanford University, 2Department of Radiology, Stanford University
The surgical procedure for delivery of embryonic stem cell-derived endothelial cells to the ischemic hindlimb is demonstrated, with non-invasive tracking by bioluminescence imaging.
Published January 23, 2009. Keywords: Medicine, hindlimb ischemia, peripheral arterial disease, embryonic stem cell, cell transplantation, bioluminescence imaging, non-invasive tracking, mouse model
1Medical Pharmacology and Physiology, University of Missouri, 2Dalton Cardiovascular Research Center
We present a preparation for visualizing and manipulating calcium signaling in native, intact microvascular endothelium. Endothelial tubes freshly isolated from mouse resistance arteries supplying skeletal muscle retain in vivo morphology and dynamic signaling within and between neighboring cells. Endothelial tubes can be prepared from microvessels of other tissues and organs.
Published November 25, 2013. Keywords: Basic Protocol, endothelial tubes, microcirculation, calcium signaling, resistance vasculature, Confocal microscopy
JoVE Immunology and Infection
1NIHR Biomedical Research Unit, Centre for Liver Research, School of Immunity and Infection, University of Birmingham
Leucocyte recruitment to the liver occurs within the specialized channels of the hepatic sinusoids which are lined by unique hepatic sinusoidal endothelial cells. Phase contrast microscopy of leucocyte recruitment across human hepatic sinusoidal endothelium under conditions of physiological shear stress can facilitate the elucidation of the molecular mechanisms which underlie this process.
Published March 21, 2014. Keywords: Immunology, Leucocyte trafficking, liver, hepatic sinusoidal endothelial cells, peripheral blood lymphocytes, flow adhesion assay
1Center for Stem Cell Research and Regenerative Medicine, Tulane University School of Medicine, 2Division of Regenerative Medicine, Tulane National Primate Research Center, 3Department of Microbiology and Immunology, Tulane University School of Medicine, 4Department of Pharmacology, Tulane University School of Medicine
Whole-organ decellularization produces natural biological scaffolds that may be used for regenerative medicine. The description of a nonhuman primate model of lung regeneration in which whole lungs are decellularized and then seeded with adult stem cells and endothelial cells in a bioreactor that facilitates vascular circulation and liquid media ventilation is presented.
Published December 15, 2013. Keywords: Bioengineering, rhesus macaque, decellularization, recellularization, detergent, matrix, scaffold, large-organ bioreactor, mesenchymal stem cells