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Assessment of Oxidative Damage in the Primary Mouse Ocular Surface Cells/Stem Cells in Response to Ultraviolet-C (UV-C) Damage
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JoVE Journal Developmental Biology
Assessment of Oxidative Damage in the Primary Mouse Ocular Surface Cells/Stem Cells in Response to Ultraviolet-C (UV-C) Damage

Assessment of Oxidative Damage in the Primary Mouse Ocular Surface Cells/Stem Cells in Response to Ultraviolet-C (UV-C) Damage

DOI:
10.3791/59924-v

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12:59 min

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February 15, 2020

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, , , , ,
1Stem Cells and Regenerative Medicine Centre,Yenepoya Research Centre, Yenepoya (Deemed to be University), Mangaluru-575018, Karnataka, India

Chapters

  • 00:00Introduction
  • 01:36Ultraviolet C (UV-C) Radiation Exposure to Mouse Primary Ocular Cells
  • 03:43Preparation of the Living-Cell Staining Media
  • 04:58Live Cell Staining of UV-C Exposed Mouse Primary Ocular Cells
  • 05:51Viewing of DCFDA (ROS), Hoechst and PI Stained Cells
  • 07:40Results
  • 11:57Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol demonstrates the simultaneous detection of reactive oxygen species (ROS), live cells, and dead cells in live primary cultures from mouse ocular surface cells. 2',7'-Dichlorofluoresceindiacetate, propidium iodide, and Hoechst staining are used to assess the ROS, dead cells, and live cells, respectively, followed by imaging and analysis.

Tags

Keywords: Oxidative DamagePrimary Mouse Ocular Surface CellsStem CellsUltraviolet-C (UV-C) DamageReactive Oxygen Species (ROS)DCFDAPropidium IodideHoechstLive Cell PermeantDead Cell PermeantUV CrosslinkerDose-response

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