Generation of an immortalized human choroid endothelial cell line (iChEC-1) using an endothelial cell specific promoter.

Abstract:

:Age-related macular degeneration (AMD) is a common cause of blindness worldwide. While recent studies have revealed that the loss of choroidal endothelial cells (ChECs) is critical to the disease pathogenesis of dry AMD, in vitro studies are needed to fully elucidate the disease mechanism. However, these studies remain hindered due to the lack of publically available human ChEC lines. To address this need, ChECs were harvested form donor tissue and enriched for by using magnetic cell separation using anti-CD31 conjugated microbeads. Next, lenti-viral vectors with endothelial-specific promoters driving genes necessary for immortalization, CDH5p-hTERT and CDH5p TAg, were generated. Stable integration of both gene cassettes allowed cells to maintain their proliferative state and yielded an immortalized cell line (iChEC-1). Immunocytochemical analysis of iChEC-1 confirmed the expression of important ChEC markers such as CA4, a marker of choriocapillaris endothelial cells, CDH5, and CD34, pan-endothelial cell markers. qRT-PCR analysis of expanded clones from iChEC-1 further showed that the line maintained expression of other important endothelial markers, vWF, PECAM1, and PLVAP, similar to primary cells. Functional responses were characterized by tube-forming assays and repopulation of decellularized choroid with the immortalized cell line. In conclusion, the iChEC-1 line presents a suitable immortalized human ChEC line for future in vitro studies of AMD.

journal_name

Microvasc Res

journal_title

Microvascular research

authors

Giacalone JC,Miller MJ,Workalemahu G,Reutzel AJ,Ochoa D,Whitmore SS,Stone EM,Tucker BA,Mullins RF

doi

10.1016/j.mvr.2018.12.002

subject

Has Abstract

pub_date

2019-05-01 00:00:00

pages

50-57

eissn

0026-2862

issn

1095-9319

pii

S0026-2862(18)30221-8

journal_volume

123

pub_type

杂志文章
  • Tissue-isolated human tumor xenografts in athymic nude mice.

    abstract::An ex vivo perfused solid tumor preparation provides control over the physiological, biochemical, and pharmacological composition of the arterial input and easy access to the venous output. This is advantageous for studies of transport and metabolism in solid tumors. Here we present a tissue-isolated tumor preparation...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1006/mvre.1994.1063

    authors: Kristjansen PE,Roberge S,Lee I,Jain RK

    更新日期:1994-11-01 00:00:00

  • Feasibility of assessment of conjunctival microvascular hemodynamics in unilateral ischemic stroke.

    abstract::Since the internal carotid artery supplies blood to both the eye and the brain, ocular microvascular hemodynamics can be altered due to ischemic stroke. The purpose of the current study was to establish the feasibility of conjunctival microcirculation imaging for detection of inter-ocular differences in microvascular ...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1016/j.mvr.2015.04.007

    authors: Kord Valeshabad A,Wanek J,Mukarram F,Zelkha R,Testai FD,Shahidi M

    更新日期:2015-07-01 00:00:00

  • Expression of miR-210 in the peripheral blood of patients with newly diagnosed type 2 diabetes mellitus and its effect on the number and function of endothelial progenitor cells.

    abstract:OBJECTIVE:This study aims to investigate the correlation between the expression of miR-210 in peripheral blood and the number of peripheral endothelial progenitor cells (EPCs) in patients with type 2 diabetes mellitus (T2DM). We also determined the effect of miR-210 on EPC proliferation, adhesion, migration, tube forma...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1016/j.mvr.2020.104032

    authors: Li X,Jia Z,Zhao X,Xu M,Chen M

    更新日期:2020-09-01 00:00:00

  • Role of histamine release in nonspecific vasodilatation during anodal and cathodal iontophoresis.

    abstract::Nonspecific vasodilatation during iontophoresis is an important confounding factor in experimental pharmacology. In this investigation, we studied the involvement of sensory nerves and histamine-related reactions in causing nonspecific vasodilatation in a model of anodal and cathodal iontophoresis of sodium chloride. ...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1016/j.mvr.2003.12.002

    authors: Horiuchi Y,Droog EJ,Henricson J,Wikström T,Lennquist S,Sjöberg F

    更新日期:2004-03-01 00:00:00

  • Strychnine inhibits inflammatory angiogenesis in mice via down regulation of VEGF, TNF-α and TGF-β.

    abstract::Strychnine is known to possess anti-inflammatory and antitumour activity, but its roles in tumour angiogenesis, the key step involved in tumour growth and metastasis, and the involved molecular mechanism are still unknown. We aimed to investigate the effects of strychnine on key components of inflammatory angiogenesis...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1016/j.mvr.2013.01.003

    authors: Saraswati S,Agarwal SS

    更新日期:2013-05-01 00:00:00

  • Polyarginine and eosinophil-derived major basic protein increase microvascular permeability independently of histamine or nitric oxide release.

    abstract::We tested the hypothesis that cationic peptides (polyarginine; poly-argn) and eosinophil-derived major basic protein (MaBP) increase permeability by stimulating the release of histamine and/or nitric oxide. We used intravital microscopy, clearance, and integrated optical intensity (IOI), using FITC-dextran 150 (FITC-d...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1006/mvre.1995.1038

    authors: Minnicozzi M,Ramírez MM,Egan RW,Gleich GJ,Kobayashi I,Kim D,Durán WN

    更新日期:1995-07-01 00:00:00

  • Intracellular localization of lipoplexed siRNA in vascular endothelial cells of different mouse tissues.

    abstract::Liposomally formulated siRNA can be used for RNAi applications in vivo. Intravenous bolus administration of lipoplexed siRNA has been shown to reduce gene expression in the vascular endothelium. Here, we applied immunofluorescence staining for different endothelial markers (PECAM-1, CD34, laminin) on paraffin sections...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1016/j.mvr.2008.02.004

    authors: Aleku M,Fisch G,Möpert K,Keil O,Arnold W,Kaufmann J,Santel A

    更新日期:2008-05-01 00:00:00

  • Transport properties of the mesothelium and interstitium measured in rabbit pericardium.

    abstract::The contribution of the pleural mesothelium to pleural liquid and protein transport is still vigorously debated. Recent in vitro studies of stripped pleural membrane and free-standing pericardium have demonstrated active ion solute coupled transport of liquid and transcytosis of protein. However, the relative contribu...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1016/j.mvr.2005.10.003

    authors: Tang SM,Lai-Fook SJ

    更新日期:2005-11-01 00:00:00

  • Mechanisms which mediate the antiapoptotic effects of angiopoietin-1 on endothelial cells.

    abstract::The main objective of this study was to identify molecular mechanisms through which angiopoietin-1 (Ang-1), a ligand for Tie-2 receptors, influences endothelial cell apoptosis. Human umbilical vein endothelial cells were cultured in a medium enriched with 2% fetal bovine serum (FBS) and growth supplements. Apoptosis w...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1006/mvre.2002.2421

    authors: Harfouche R,Hasséssian HM,Guo Y,Faivre V,Srikant CB,Yancopoulos GD,Hussain SN

    更新日期:2002-07-01 00:00:00

  • Automated segmentation and fractal analysis of high-resolution non-invasive capillary perfusion maps of the human retina.

    abstract::The retina provides a window to study the pathophysiology of cerebrovascular diseases. Pathological retinal microvascular changes may reflect microangiopathic processes in the brain. Recent advances in optical imaging techniques have enabled the imaging of the retinal microvasculature at the capillary level, and the g...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1016/j.mvr.2013.06.008

    authors: Jiang H,Debuc DC,Rundek T,Lam BL,Wright CB,Shen M,Tao A,Wang J

    更新日期:2013-09-01 00:00:00

  • Infrared imaging of nitric oxide-mediated blood flow in human sickle cell disease.

    abstract::Vascular dysfunction is an important pathophysiologic manifestation of sickle cell disease (SCD), a condition that increases risk of pulmonary hypertension and stroke. We hypothesized that infrared (IR) imaging would detect changes in cutaneous bloodflow reflective of vascular function. We performed IR imaging and con...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1016/j.mvr.2012.06.011

    authors: Gorbach AM,Ackerman HC,Liu WM,Meyer JM,Littel PL,Seamon C,Footman E,Chi A,Zorca S,Krajewski ML,Cuttica MJ,Machado RF,Cannon RO 3rd,Kato GJ

    更新日期:2012-11-01 00:00:00

  • Fibroblasts are in a position to provide directional information to migrating neutrophils during pneumonia in rabbit lungs.

    abstract::Previous findings have shown that pulmonary fibroblasts are associated with preexisting holes in the endothelial and epithelial basal laminae through which neutrophils appear to enter and leave the interstitium as they migrate from capillaries to alveoli. To determine their role in neutrophil migration, fibroblast org...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1006/mvre.1996.0029

    authors: Behzad AR,Chu F,Walker DC

    更新日期:1996-05-01 00:00:00

  • Pulsed acoustic cellular expression as a protective therapy against I/R injury in a cremaster muscle flap model.

    abstract:BACKGROUND:Tissue ischemia and reperfusion (I/R) affects blood flow restoration and oxygen delivery to the damaged tissues contributing to tissue morbidity and microcirculatory compromise. Pulsed acoustic cellular expression (PACE) technology is known to support tissue neovascularization. The aim of this study was to t...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1016/j.mvr.2011.11.005

    authors: Krokowicz L,Klimczak A,Cwykiel J,Mielniczuk M,Grykien C,Siemionow M

    更新日期:2012-03-01 00:00:00

  • Effects of endotoxin on lung pericytes in vitro.

    abstract::Lipopolysaccharide released during bacterial sepsis causes acute lung injury and ARDS. Pulmonary microvascular injury is a feature of ARDS, and vascular remodeling develops, leading to pulmonary hypertension. Pericytes in the lung circulation proliferate and contribute to the remodeling seen in experimental sepsis. It...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1006/mvre.1998.2085

    authors: Khoury J,Langleben D

    更新日期:1998-09-01 00:00:00

  • Altered regulator mechanisms of presynaptic adrenergic nerve: a new physiopathological hypothesis in Raynaud's disease.

    abstract::The effect of isoprenaline infusion (1 gamma/min) on blood flow in the extremities was studied in 15 Raynaud's disease (RD) patients and 9 healthy controls. A highly significant (P less than 0.0005) decrease in blood flow was seen in the RD subjects. It is suggested that an increased presynaptic beta receptor response...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1016/0026-2862(84)90045-1

    authors: Giovanni B,Giuseppina CM,Susanna F,Roberto M

    更新日期:1984-01-01 00:00:00

  • Effect of diffusion boundary layers on the initial uptake of O2 by red cells. Theory versus experiment.

    abstract::We have applied theories of mass transfer, in laminar and turbulent flow, to red cells in the stopped-flow apparatus in order to estimate the effect of extracellular diffusion boundary layers on the initial rate of O2 uptake. We compared the theoretical predictions with the results of our previous stopped-flow experim...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1016/0026-2862(83)90058-4

    authors: Huxley VH,Kutchai H

    更新日期:1983-07-01 00:00:00

  • Isolation, culture, and characterisation of MHC class II-positive microvascular endothelial cells from the human heart.

    abstract::Immunological studies of human endothelial cells have been performed almost entirely on endothelial cells (EC) derived from foetal (umbilical vein) or adult (aorta) large vessels. Immunologically these EC have in common the fact that they are negative for major histocompatibility complex (MHC) class II antigens (HLA-D...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1006/mvre.1996.0016

    authors: McDouall RM,Yacoub M,Rose ML

    更新日期:1996-03-01 00:00:00

  • Sanguinate's effect on pial arterioles in healthy rats and cerebral oxygen tension after controlled cortical impact.

    abstract::Sanguinate, a polyethylene glycol-conjugated carboxyhemoglobin, was investigated for cerebral vasoactivity in healthy male Sprague-Dawley rats (Study 1) and for its ability to increase brain tissue oxygen pressure (PbtO2) after controlled cortical impact (CCI) - traumatic brain injury (TBI) (Study 2). In both studies ...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1016/j.mvr.2016.06.001

    authors: Mullah SH,Abutarboush R,Moon-Massat PF,Saha BK,Haque A,Walker PB,Auker CR,Arnaud FG,McCarron RM,Scultetus AH

    更新日期:2016-09-01 00:00:00

  • Role of prostanoids and 20-HETE in mediating oxygen-induced constriction of skeletal muscle resistance arteries.

    abstract::This study determined the contribution of cytochrome P450 (CP450) 4A enzyme metabolites of arachidonic acid in mediating the constriction of isolated rat skeletal muscle resistance arteries in response to elevated PO2. Gracilis arteries (GA) were viewed via television microscopy and constrictor responses to elevated P...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1006/mvre.2001.2341

    authors: Frisbee JC,Krishna UM,Falck JR,Lombard JH

    更新日期:2001-11-01 00:00:00

  • A new view of Starling's hypothesis at the microstructural level.

    abstract::In this paper we quantitatively investigate the hypothesis proposed by Michel (Exp. Physiol. 82, 1-30, 1997) and Weinbaum (Ann. Biomed. Eng. 26, 1-17, 1998) that the Starling forces are determined by the local difference in the hydrostatic and colloid osmotic pressure across the endothelial surface glycocalyx, which w...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1006/mvre.1999.2177

    authors: Hu X,Weinbaum S

    更新日期:1999-11-01 00:00:00

  • The effect of treatment with low dose ACE inhibitor and/or diuretic on coronary microvasculature in stroke-prone spontaneously hypertensive rats.

    abstract::Angiotensin II is considered to have angiogenic properties. Nevertheless, several authors reported an increase in coronary capillary density after treatment with ACE inhibitors. The aim of the present study was to evaluate the effect of treatment with low doses of ACE inhibitor perindopril, low doses of the diuretic i...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1006/mvre.1999.2224

    authors: Rakusan K,Cicutti N,Maurin A,Guez D,Schiavi P

    更新日期:2000-03-01 00:00:00

  • Lymphangiogenesis following obstruction of large postnodal lymphatics in sheep.

    abstract::We examined the impact of lymph flow obstruction in large post-nodal lymphatic vessels in sheep. A silk ligature was placed 2 cm downstream from the prescapular or popliteal lymph node and tightened to interrupt flow. At 6, 12 and 16 weeks after lymph flow blockage, a network of small interconnecting lymphatics (appro...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1016/j.mvr.2006.11.003

    authors: Jila A,Kim H,Nguyen VP,Dumont DJ,Semple J,Armstrong D,Seto E,Johnston M

    更新日期:2007-05-01 00:00:00

  • Adenosine enhances functional activation of blood flow in cat optic nerve head during photic stimulation independently from nitric oxide.

    abstract::Blood flow studies in the brain, heart, and other organs suggest that there could be interaction between nitric oxide (NO) and adenosine. This possibility was investigated in the optic nerve head (ONH) during photic stimulation of the dark-adapted cat eye. Functional activation of ONH blood flow was measured by laser ...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1006/mvre.2002.2432

    authors: Buerk DG,Riva CE

    更新日期:2002-09-01 00:00:00

  • Chronic elevations in salt intake and reduced renal mass hypertension compromise mechanisms of arteriolar dilation.

    abstract::Alterations in the reactivity of arterioles to vasodilator agonists were assessed in the skeletal muscle microcirculation of age-matched, normotensive male Sprague-Dawley rats fed either a high salt (4% NaCl, HS) or a low salt (0.4% NaCl, LS) diet and in reduced renal mass hypertensive rats on a high salt diet (HSRRM)...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1006/mvre.1998.2105

    authors: Frisbee JC,Lombard JH

    更新日期:1998-11-01 00:00:00

  • Endothelial progenitor cells--an evolving story.

    abstract::The first description of endothelial progenitor cells (EPC) in 1997 led rapidly to substantial changes in our understanding of angiogenesis, and within 5 years to the first clinical studies in humans using bone marrow derived EPC to enhance coronary neovascularisation and cardiac function after myocardial ischemia. Ho...

    journal_title:Microvascular research

    pub_type: 杂志文章,评审

    doi:10.1016/j.mvr.2009.12.004

    authors: Pearson JD

    更新日期:2010-05-01 00:00:00

  • Contractile responses of isolated rat mesenteric arteries to acute episodes of severe hypoxia and subsequent reoxygenation.

    abstract::This study further investigates the mechanisms responsible for the effects of acute and severe hypoxia, and subsequent reoxygenation, on the contractility of isolated rat mesenteric arteries. In noradrenaline (NA)-contracted arteries, hypoxia caused a relaxation to near baseline levels. Reoxygenation resulted in an im...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1016/j.mvr.2004.08.001

    authors: Bruce J,Taggart M,Austin C

    更新日期:2004-11-01 00:00:00

  • Modification of vascular and inflammation biomarkers after OGTT in overweight healthy and diabetic subjects.

    abstract::We evaluated the effect of an oral glucose tolerance test (OGTT) on the level of biomarkers of vascular remodelling. We enrolled 256 Caucasian overweight healthy subjects (H) and 274 overweight type 2 diabetic patients (D). All patients underwent basal measurements of blood glucose (BG), nitrites/nitrates, adiponectin...

    journal_title:Microvascular research

    pub_type: 杂志文章,多中心研究

    doi:10.1016/j.mvr.2010.01.002

    authors: Derosa G,D'Angelo A,Salvadeo SA,Ferrari I,Fogari E,Gravina A,Mereu R,Palumbo I,Maffioli P,Randazzo S,Cicero AF

    更新日期:2010-03-01 00:00:00

  • Effect of concentration and hydration on restriction of albumin by lung interstitium.

    abstract::In previous studies, the flow of albumin solution through hydrated lung interstitial segments was higher than a prior flow of Ringer solution (A. Tajaddinni et al., 1994, J. Appl. Physiol. 76, 578-583). We wondered whether this effect was caused by an increased pore size. We measured the flow of albumin solutions thro...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1006/mvre.2001.2363

    authors: Villarruel S,Ibbott GS,Lai-Fook SJ

    更新日期:2002-01-01 00:00:00

  • Acute VEGF effect on solute permeability of mammalian microvessels in vivo.

    abstract::To investigate the effect of vascular endothelial growth factor (VEGF) on solute permeability of mammalian microvessels, we measured the apparent permeability (P) of various-sized solutes on the postcapillary venules of rat mesentery in vivo. Exposure to 1 nM VEGF transiently increased P from a mean of 1.4 (+/-0.11 SE...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1016/j.mvr.2004.03.004

    authors: Fu BM,Shen S

    更新日期:2004-07-01 00:00:00

  • Endothelial NO/cGMP system contributes to natriuretic peptide-mediated coronary and peripheral vasodilation.

    abstract::We tested the hypothesis that the endothelial nitric oxide (NO)-soluble guanylyl cyclase system is involved in atrial natriuretic peptide (ANP) and C-type natriuretic peptide (CNP) mediated regulation of coronary and peripheral vascular resistance. Rat hearts were perfused via the aorta at constant flow and the effect...

    journal_title:Microvascular research

    pub_type: 杂志文章

    doi:10.1006/mvre.2000.2289

    authors: Brunner F,Wölkart G

    更新日期:2001-01-01 00:00:00