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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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Biologia do Desenvolvimento
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JoVE Journal Biologia do Desenvolvimento
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

Capítulos

  • 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
  • 07:40Resultados
  • 11:57Conclusion

Summary

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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

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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