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Isolation of Murine Retinal Endothelial Cells for Next-Generation Sequencing
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Developmental Biology
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JoVE Journal Developmental Biology
Isolation of Murine Retinal Endothelial Cells for Next-Generation Sequencing

Isolation of Murine Retinal Endothelial Cells for Next-Generation Sequencing

DOI:
10.3791/63133-v

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09:59 min

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October 11, 2021

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, , ,
1Department of Cell Biology,University of Virginia School of Medicine, 2Robert M. Berne Cardiovascular Research Center,University of Virginia School of Medicine, 3Department of Cardiology,University of Virginia School of Medicine, 4Hematovascular Biology Center,University of Virginia School of Medicine, 5Yale Cardiovascular Research Center,Yale University School of Medicine

Chapters

  • 00:04Introduction
  • 00:38Obtain Mouse Eyes for Retinal Isolation and Isolate Mouse Retinal Tissue
  • 02:30Digest Retinal Tissue into a Single Cell Suspension and Immunostain the Cells with Antibodies
  • 04:07Prepare for Fluorescence‐Activated Cell Sorting
  • 05:19Isolate Viable Endothelial Cells via FACS
  • 07:18Results: Analysis of Isolated Endothelial Cells and Next‐Generation Sequencing After Isolation
  • 09:03Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol describes a method for the isolation of murine postnatal retinal endothelial cells optimized for cell yield, purity, and viability. These cells are suitable for next-generation sequencing approaches.

Tags

Keywords: Retinal Endothelial CellsNext-generation SequencingVascular DevelopmentNeonatal MouseEye DissectionCell Isolation

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