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Analysis of Retinoic Acid-induced Neural Differentiation of Mouse Embryonic Stem Cells in Two and Three-dimensional Embryoid Bodies
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Biologia dello sviluppo
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JoVE Journal Biologia dello sviluppo
Analysis of Retinoic Acid-induced Neural Differentiation of Mouse Embryonic Stem Cells in Two and Three-dimensional Embryoid Bodies

Analysis of Retinoic Acid-induced Neural Differentiation of Mouse Embryonic Stem Cells in Two and Three-dimensional Embryoid Bodies

DOI:
10.3791/55621-v

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09:04 min

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April 22, 2017

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, , ,
1Department of Medicine, Cardeza Vascular Research Center,Sidney Kimmel Medical College, Thomas Jefferson University, 2Department of Molecular Cardiology,Cleveland Clinic Foundation, 3Department of Cancer Biology, Cardeza Vascular Research Center,Sidney Kimmel Medical College, Thomas Jefferson University

Capitoli

  • 00:05Titolo
  • 00:51Embryoid Body Formation
  • 01:582D Embryoid Body Culture and Detection of Secreted Proteins
  • 03:043D Embryoid Body Culture
  • 04:06Embryoid Body Dissociation
  • 05:56Analysis of Embryoid Body Formation by Immunofluorescence
  • 07:12Results: Characteristics of Embryo Body Cultures and Visualization of Neural Differentiation Markers
  • 08:22Conclusion

Summary

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We describe a technique for using murine embryonic stem cells for generating two or three dimensional embryoid bodies. We then explain how to induce neural differentiation of the embryoid body cells by retinoic acid, and how to analyze their state of differentiation by progenitor cell marker immunofluorescence and immunoblotting.

Tags

Keywords: Embryonic Stem CellsNeural DifferentiationRetinoic AcidEmbryoid BodiesGene ExpressionProtein MarkersOptical MicroscopyTrypsin-EDTAIMDMFBSHanging DropsFibronectin-coated CoverslipsProtein Secretion AnalysisCentrifugal Filters

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