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In Vitro Differentiation of Mouse Granulocyte-macrophage-colony-stimulating Factor (GM-CSF)-producing T Helper (THGM) Cells
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JoVE Journal Immunology and Infection
In Vitro Differentiation of Mouse Granulocyte-macrophage-colony-stimulating Factor (GM-CSF)-producing T Helper (THGM) Cells

In Vitro Differentiation of Mouse Granulocyte-macrophage-colony-stimulating Factor (GM-CSF)-producing T Helper (THGM) Cells

DOI:
10.3791/58087-v

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10:27 min

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September 10, 2018

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, ,
1Department of Microbiology and Immunology, Yong Loo Lin School of Medicine,National University of Singapore, 2Immunology Programme, Life Sciences Institute,National University of Singapore, 3Cancer Science Institute of Singapore, Yong Loo Lin School of Medicine,National University of Singapore

Chapters

  • 00:04Title
  • 01:01Murine Splenocyte Preparation
  • 02:10CD4+ T Cell Magnetic Bead Separation
  • 03:48Naïve (CD25-CD44loCD62Lhi) CD4+ T Cell Fluorescence-activated Cell Sorting (FACS) and Granulocyte Macrophage-colony-stimulating Factor (GM-CSF) T Helper (THGM) Cell Differentiation
  • 05:48Mouse THGM Cell Analysis
  • 07:35Results: THGM Cytokine Profile Analysis
  • 09:30Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we present a protocol to differentiate murine granulocyte-macrophage-colony-stimulating-factor-producing T helper (THGM) cells from naive CD4+ T cells, including isolation of naive CD4+ T cells, differentiation of THGM, and analysis of differentiated THGM cells. This method can be applied to studies of the regulation and function of THGM cells.

Tags

Keywords: CD4 T-cellsGM-CSFT Helper CellsTHGM CellsIn Vitro DifferentiationInflammatory DiseasesMultiple SclerosisCell IsolationCell CultureMagnetic Separation

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