Methods Article

Isolation of Murine Retinal Endothelial Cells for Next-Generation Sequencing

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2.9K Views

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09:59 min

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October 11th, 2021

DOI :

10.3791/63133-v

October 11th, 2021

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Nicholas W. Chavkin¹^,², Shelby Cain¹, Kenneth Walsh²^,³^,⁴, Karen K. Hirschi¹^,²^,⁴^,⁵

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

Podsumowanie

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Murine Retinal Endothelial Cells

Isolation Protocol

Next generation Sequencing

Vascular Development

Cell Viability

Laboratory Techniques

Dissection Microscope

Neonatal Mouse

PBS Solution

Retinal Tissue Preparation

Digestion Solution

Pipetting Technique

Tissue Extraction

Micro centrifuge Tubes

Rozdziały w tym wideo

0:04

Introduction

0:38

Obtain Mouse Eyes for Retinal Isolation and Isolate Mouse Retinal Tissue

2:30

Digest Retinal Tissue into a Single Cell Suspension and Immunostain the Cells with Antibodies

4:07

Prepare for Fluorescence‐Activated Cell Sorting

5:19

Isolate Viable Endothelial Cells via FACS

7:18

Results: Analysis of Isolated Endothelial Cells and Next‐Generation Sequencing After Isolation

9:03

Conclusion

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