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Co-Culture and Transduction of Murine Thymocytes on Delta-Like 4-Expressing Stromal Cells to Study Oncogenes in T-Cell Leukemia
JoVE Journal
Cancer Research
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JoVE Journal Cancer Research
Co-Culture and Transduction of Murine Thymocytes on Delta-Like 4-Expressing Stromal Cells to Study Oncogenes in T-Cell Leukemia

Co-Culture and Transduction of Murine Thymocytes on Delta-Like 4-Expressing Stromal Cells to Study Oncogenes in T-Cell Leukemia

DOI:
10.3791/64271-v

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09:43 min

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June 09, 2023

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, , , , , , ,
1Cytokines and Immunity Section, Cancer Innovation Laboratory, National Cancer Institute,National Institutes of Health, 2Comparative Biomedical Scientist Training Program,NIH, 3Department of Pathobiology and Diagnostic Investigation, Veterinary Diagnostic Laboratory,Michigan State University, 4NCI-Frederick Laboratory Animal Sciences Program

Chapters

  • 00:04Introduction
  • 00:47Seeding and Transfecting the Retrovirus-Producer Cells
  • 02:31Thymocyte Preparation and Depletion of CD4+ and CD8+ Cells
  • 04:53Harvesting Retrovirus and Transducing the Thymocytes
  • 07:13Results: Thymocyte Depletion and Thymocyte Retroviral Transduction Efficiency
  • 08:51Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol describes the isolation of double-negative thymocytes from the mouse thymus followed by retroviral transduction and co-culture on the delta-like 4-expressing bone marrow stromal cell line co-culture system (OP9-DL4) for further functional analysis.

Tags

Co-cultureTransductionMurine ThymocytesDelta-Like 4Stromal CellsT-cell LeukemiaRetroviral VectorsOP9 DL4In Vitro SystemT-cell DevelopmentCancerThymocyte SuspensionThymusPBSGlass SlidesCell Filtration

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