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Study of Dendritic Cell Development by Short Hairpin RNA-Mediated Gene Knockdown in a Hematopoietic Stem and Progenitor Cell Line In vitro
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JoVE Journal Immunologia e infezioni
Study of Dendritic Cell Development by Short Hairpin RNA-Mediated Gene Knockdown in a Hematopoietic Stem and Progenitor Cell Line In vitro

Study of Dendritic Cell Development by Short Hairpin RNA-Mediated Gene Knockdown in a Hematopoietic Stem and Progenitor Cell Line In vitro

DOI:
10.3791/62730-v

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06:12 min

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March 07, 2022

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, ,
1Graduate Institute of Immunology,National Taiwan University College of Medicine, 2Department of Pathology, Division of Microbiology and Immunology,University of Utah

Capitoli

  • 00:04Introduction
  • 00:43Preparation of Immortalized HematopoieticStem and Progenitor Cell (iHSPC) Lines and Lentiviral Transduction
  • 02:08In Vitro Differentiation of the Stable Knockdown iHSPC Cell Lines
  • 03:05Flow Cytometric Analysis of the Differentiated Dendritic Cells
  • 04:01Results: Dendritic Cell Differentiation Following Short Hairpin RNA-Mediated Tcf4 and Id2 Knockdown
  • 05:33Conclusion

Summary

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Here we provide a protocol for screening potential transcription factors involved in the development of dendritic cell (DC) using lentiviral transduction of shRNA to obtain stable knockdown cell lines for in vitro DC differentiation.

Tags

Keywords: Dendritic Cell DevelopmentShort Hairpin RNAGene KnockdownHematopoietic Stem And Progenitor Cell LineIn VitroTranscription FactorsFLT3 LigandBeta EstradiolLentiviral TransductionPuromycinLacZTcf4Id2Flow Cytometry

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