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Retroviral Infection of Murine Embryonic Stem Cell Derived Embryoid Body Cells for Analysis of Hematopoietic Differentiation
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Retroviral Infection of Murine Embryonic Stem Cell Derived Embryoid Body Cells for Analysis of Hematopoietic Differentiation

Retroviral Infection of Murine Embryonic Stem Cell Derived Embryoid Body Cells for Analysis of Hematopoietic Differentiation

DOI:
10.3791/52022-v

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11:40 min

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October 20, 2014

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, , ,
1Harper Cancer Research Institute, 2Microbiology and Immunology,Indiana University School of Medicine, 3Department of Biological Sciences,University of Notre Dame

Chapters

  • 00:05Title
  • 01:58Virus Infection of Embryoid Body (EB) Cells
  • 04:02Differentiating Viral Infected EB Cells in Hanging Drops
  • 05:47Preparing Single Cell Suspensions for Flow Cytometer Analysis
  • 07:02Flow Cytometry Analysis for Hematopoietic Progenitors
  • 08:48Results: Analysis of Retroviral Infected EBs and Hematopoietic Progenitor Cell Generation
  • 11:00Conclusion

Summary

Automatic Translation

Automatic Translation

Manipulating temporal gene expression in differentiating embryonic stem cells (ESCs) can be achieved using inducible gene systems. However, generation of these cell lines is costly and time consuming. This protocol achieves rapid expression of a transgene in differentiating ES-derived cells and subsequent analysis of downstream hematopoietic differentiation.

Tags

Keywords: Embryonic Stem CellsHematopoietic Progenitor CellsEmbryoid BodiesRetroviral InfectionGene ExpressionInducible Transgene SystemFlow CytometryMicroRNAMesodermHematopoietic Differentiation

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