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Gene Expression Analysis of Endothelial Cells Exposed to Shear Stress Using Multiple Parallel-plate Flow Chambers
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Bioengineering
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JoVE Journal Bioengineering
Gene Expression Analysis of Endothelial Cells Exposed to Shear Stress Using Multiple Parallel-plate Flow Chambers

Gene Expression Analysis of Endothelial Cells Exposed to Shear Stress Using Multiple Parallel-plate Flow Chambers

DOI:
10.3791/58478-v

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08:50 min

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October 21, 2018

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, , , , ,
1Institute of Medical Science,University of Toronto, 2Keenan Research Centre in the Li Ka Shing Knowledge Institute,St. Michael’s Hospital, 3Department of Laboratory Medicine and Pathobiology,University of Toronto, 4Department of Medicine,University of Toronto

Chapters

  • 00:04Title
  • 00:55Assay Setup
  • 04:20Flow Assay
  • 05:18Cell Harvest and RNA Extraction
  • 06:48Results: Representative KLF2 Loss-of-Function Analysis in Human Endothelial Cells in Response to Shear Stress
  • 08:06Conclusion

Summary

Automatic Translation

Automatic Translation

Here, a workflow for the culture and gene expression analysis of endothelial cells under fluid shear stress is presented. Included is a physical arrangement for simultaneously housing and monitoring multiple flow chambers in a controlled environment and the use of an exogenous reference RNA for quantitative PCR.

Tags

Keywords: Endothelial CellsShear StressParallel-plate Flow ChambersGene Expression AnalysisAtherosclerosisLaminar FlowCell CultureFibronectinCO2 IncubatorBubble TrapCell SeedingTubingStopcock

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