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In Vitro Three-Dimensional Sprouting Assay of Angiogenesis Using Mouse Embryonic Stem Cells for Vascular Disease Modeling and Drug Testing
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Biologia
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JoVE Journal Biologia
In Vitro Three-Dimensional Sprouting Assay of Angiogenesis Using Mouse Embryonic Stem Cells for Vascular Disease Modeling and Drug Testing

In Vitro Three-Dimensional Sprouting Assay of Angiogenesis Using Mouse Embryonic Stem Cells for Vascular Disease Modeling and Drug Testing

DOI:
10.3791/62554-v

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08:04 min

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May 11, 2021

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, , ,
1Einthoven Laboratory for Experimental Vascular Medicine, Department of Internal Medicine (Nephrology),Leiden University Medical Center, 2Institute Physics for Medicine Paris,INSERM U1273, ESPCI Paris, CNRS FRE 2031, 3MEMOLIFE Laboratory of Excellence and PSL Research University

Capitoli

  • 00:04Introduction
  • 01:14Embryoid Bodies (EB) Formation in Hanging Drop
  • 03:02Floating EBs Culture for Vascular Differentiation
  • 03:413D Sprouting Angiogenesis Assay and Immunofluorescence Staining
  • 05:44Results: Microscopic Analysis and Quantification of Mouse EBs Undergoing Angiogenesis Assay
  • 07:29Conclusion

Summary

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This assay utilizes mouse embryonic stem cells differentiated into embryoid bodies cultured in 3D-collagen gel to analyze the biological processes that control sprouting angiogenesis in vitro. The technique can be applied for testing drugs, modeling diseases, and for studying specific genes in the context of deletions that are embryonically lethal.

Tags

In VitroSprouting AngiogenesisMouse Embryonic Stem CellsVascular Disease ModelingDrug TestingEndothelial Tip CellsCell MigrationGenetic ScreeningPluripotencyKaryotypingMesoderm DifferentiationLow Attachment Polystyrene Dishes

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