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Development and Characterization of In Vitro Microvessel Network and Quantitative Measurements of Endothelial [Ca2+]i and Nitric Oxide Production
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Biologia
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JoVE Journal Biologia
Development and Characterization of In Vitro Microvessel Network and Quantitative Measurements of Endothelial [Ca2+]i and Nitric Oxide Production

Development and Characterization of In Vitro Microvessel Network and Quantitative Measurements of Endothelial [Ca2+]i and Nitric Oxide Production

DOI:
10.3791/54014-v

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09:39 min

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May 19, 2016

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, , ,
1Department of Cellular and Molecular Physiology,College of Medicine, Penn State University, 2Lane Department of Computer Science and Electrical Engineering,West Virginia University

Capitoli

  • 00:05Titolo
  • 00:58Preparing the Microchannel Device for Cell Seeding
  • 02:37Endothelial Cell Seeding and Long-term Perfusion
  • 04:38Immunofluorescent Staining and Imaging Calcium Concentration or Nitric Oxide Production
  • 06:39Data Analysis
  • 07:35Results: Endothelial Cell Culture Under Shear Stress
  • 08:39Conclusion

Summary

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Primary human umbilical vein endothelial cells (HUVECs) were grown to confluence within a microfluidic network device. The endothelial cell junction and F-actin distributions were illustrated and the changes in intracellular calcium concentration and nitric oxide production in response to adenosine triphosphate (ATP) were quantified in real-time at individual cell levels.

Tags

Keywords: In Vitro MicrovesselMicrofluidic DeviceEndothelial CellsShear FlowCalciumNitric OxidePhotolithographyPDMS MicrochannelFibronectinHUVECsDextranCell Seeding

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