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Induction and Analysis of Oxidative Stress in Sleeping Beauty Transposon-Transfected Human Retinal Pigment Epithelial Cells
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JoVE Journal Medicine
Induction and Analysis of Oxidative Stress in Sleeping Beauty Transposon-Transfected Human Retinal Pigment Epithelial Cells

Induction and Analysis of Oxidative Stress in Sleeping Beauty Transposon-Transfected Human Retinal Pigment Epithelial Cells

DOI:
10.3791/61957-v

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10:00 min

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December 11, 2020

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, , , , ,
1Experimental Ophthalmology,University of Geneva, 2Department of Ophthalmology,University Hospitals of Geneva, 3Max Delbrück Center for Molecular Medicine

Chapters

  • 00:04Introduction
  • 00:52Conditioned Medium Preparation and Collection
  • 02:47Treatment of RPE Cells with Growth Factors and H2O2
  • 04:33Glutathione Assay
  • 06:00Cytotoxicity Assay and Microscopic Analysis
  • 07:24Results: Representative Oxidative Stress Induction and Analysis
  • 09:10Conclusion

Summary

Automatic Translation

Automatic Translation

We present a protocol for the development and use ofan oxidative stress-model by treating retinal pigment epithelial cells with H2O2, analyzing cell morphology, viability, density, glutathione, and UCP-2 level. It is a useful model to investigate the antioxidant effect of proteins secreted by transposon-transfected cells to treat neuroretinal degeneration.

Tags

Keywords: Oxidative StressHydrogen PeroxideSleeping Beauty TransposonHuman Retinal Pigment Epithelial CellsAge-related Macular DegenerationNeurodegenerative DiseasesARPE-19 Cell LinePEDFGM-CSFNickel NTAProtein PurificationBCA Protein AssayConditioned MediumHydrogen Peroxide Treatment

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