Endothelial Cell Tube Formation Assay for the &lt;em&gt;In Vitro&lt;/em&gt; Study of Angiogenesis

DeCicco-Skinner, Kathleen; Henry, Gervaise H.; Cataisson, Christophe; Tabib, Tracy; Gwilliam, J. Curtis; Watson, Nicholas; Bullwinkle, Erica M.; Falkenburg, Lauren; O’Neill, Rebecca; Morin, Adam; Wiest, Jonathan S.

doi:10.3791/51312

Cited by 272 publications

(258 citation statements)

References 14 publications

Supporting

Mentioning

238

Contrasting

Unclassified

Order By: Relevance

“…Caspase-1 Inhibition Improves Tube Formation of LPCtreated Human Aortic Endothelial Cells-Because EC tube formation assay has been widely used in determining VEGFR-2-mediated angiogenesis (27), we used the EC tube formation assay in the Matrigel to test the hypothesis that caspase-1 inhibition improves VEGFR-2-mediated HAEC tube formation. To test this hypothesis, we performed tube formation assays with untreated HAECs, LPC-treated HAECs, and LPC plus caspase-1 inhibitor co-treated HAECs.…”

Section: Vegfr-2 Inhibition Increases Caspase-1 Activation In Haecsmentioning

confidence: 99%

Inhibition of Caspase-1 Activation in Endothelial Cells Improves Angiogenesis

Lopez-Pastraña

Ferrer

et al. 2015

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background:The interplay between dyslipidemia-induced inflammation and angiogenesis remains poorly understood. Results: Inhibition of caspase-1 improves VEGFR-2 signaling, tube formation, and blood perfusion in ischemic tissues. Conclusion:The suppression of caspase-1 improves angiogenesis and ischemia prognosis. Significance: Caspase-1 suppression is a novel therapeutic target for improvement of angiogenesis and ischemia under inflammatory environments.

show abstract

Section: Vegfr-2 Inhibition Increases Caspase-1 Activation In Haecsmentioning

confidence: 99%

Inhibition of Caspase-1 Activation in Endothelial Cells Improves Angiogenesis

Lopez-Pastraña

Ferrer

et al. 2015

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…EC tube formation was performed as described [24]. Briefly, HCAECs (4×10 5 /ml) were seeded in 96-well plates coated with Matrigel TM and incubated in the CM collected.…”

Section: Ec Tube Formation Assaymentioning

confidence: 99%

“…It is known that PI3K/ Akt, p38 MAPK, ERK1/2, and eNOS signaling regulate VEGF expression in ECs, endothelial progenitor cells (EPCs) and mesenchymal stem cells (MSCs) [24,25,28,32]. To test if any of these signaling pathways is involved in VNS-induced VEGF-A/B expression, we used pathway-specific inhibitors to block the above-mentioned signal individually and observe if Ach-induced VEGF expression can be altered.…”

Section: Sp1 Transcription Factor Involved In Vns-induced Vegf-a/b Exmentioning

confidence: 99%

VEGF-A and VEGF-B Coordinate the Arteriogenesis to Repair the Infarcted Heart with Vagus Nerve Stimulation

Zhong

Tang

et al. 2018

Cell Physiol Biochem

View full text Add to dashboard Cite

Background/Aims: Vagus nerve stimulation (VNS) suppresses arrhythmic activity and minimizes cardiomyocyte injury. However, how VNS affects angiogenesis/arteriogenesis in infarcted hearts, is poorly understood. Methods: Myocardial infarction (MI) was achieved by ligation of the left anterior descending coronary artery (LAD) in rats. 7 days after LAD, stainless-steel wires were looped around the left and right vagal nerve in the neck for vagus nerve stimulation (VNS). The vagal nerve was stimulated with regular pulses of 0.2ms duration at 20 Hz for 10 seconds every minute for 4 hours, and then ACh levels by ELISA in cardiac tissue and serum were evaluated for its release after VNS. Three and 14 days after VNS, Real-time PCR, immunostaining and western blot were respectively used to determine VEGF-A/B expressions and α-SMA- and CD31-postive vessels in VNS-hearts with pretreatment of α7-nAChR blocker mecamylamine (10 mg/kg, ip) or mACh-R blocker atropine (10 mg/kg, ip) for 1 hour. The coronary function and left ventricular performance were analyzed by Langendorff system and hemodynamic parameters in VNS-hearts with pretreatment of VEGF-A/B-knockdown or VEGFR blocker AMG706. Coronary arterial endothelial cells proliferation, migration and tube formation were evaluated for angiogenesis following the stimulation of VNS in coronary arterial smooth muscle cells (VSMCs). Results: VNS has been shown to stimulate VEGF-A and VEGF-B expressions in coronary arterial smooth muscle cells (VSMCs) and endothelial cells (ECs) with an increase of α-SMA- and CD31-postive vessel number in infarcted hearts. The VNS-induced VEGF-A/B expressions and angiogenesis were abolished by m-AChR inhibitor atropine and α7-nAChR blocker mecamylamine in vivo. Interestingly, knockdown of VEGF-A by shRNA mainly reduced VNS-mediated formation of CD31⁺ microvessels. In contrast, knockdown of VEGF-B powerfully abrogated VNS-induced formation of α-SMA⁺ vessels. Consistently, VNS-induced VEGF-A showed a greater effect on EC tube formation as compared to VNS-induced VEGF-B. Moreover, VEGF-A promoted EC proliferation and VSMC migration while VEGF-B induced VSMC proliferation and EC migration in vitro. Mechanistically, vagal neurotransmitter acetylcholine stimulated VEGF-A/B expressions through m/nACh-R/PI3K/Akt/Sp1 pathway in EC. Functionally, VNS improved the coronary function and left ventricular performance. However, blockade of VEGF receptor by antagonist AMG706 or knockdown of VEGF-A or VEGF-B by shRNA significantly diminished the beneficial effects of VNS on ventricular performance. Conclusion: VNS promoted angiogenesis/arteriogenesis to repair the infracted heart through the synergistic effects of VEGF-A and VEGF-B.

show abstract

“…Tubes can be visualized using a phase contrast inverted microscope. The number of branches, loops/meshes, or tubes length can be easily quantified as a measure of in vitro angiogenesis [13]. Ex vivoanti-angiogenesis assay first conducted usingrat aortic ring assay [14].It is an ex vivo physiologically relevant assay with obvious advantages over other in vitro methods.…”

Section: Introductionmentioning

confidence: 99%