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Conditional Knockdown of Gene Expression in Cancer Cell Lines to Study the Recruitment of Monocytes/Macrophages to the Tumor Microenvironment
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Cancer Research
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JoVE Journal Cancer Research
Conditional Knockdown of Gene Expression in Cancer Cell Lines to Study the Recruitment of Monocytes/Macrophages to the Tumor Microenvironment

Conditional Knockdown of Gene Expression in Cancer Cell Lines to Study the Recruitment of Monocytes/Macrophages to the Tumor Microenvironment

DOI:
10.3791/56333-v

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10:59 min

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November 23, 2017

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1Division of Hematology, Oncology and Blood and Marrow Transplantation, Department of Pediatrics,University of Southern California, 2The Saban Research Institute of Children’s Hospital Los Angeles, 3Department of Biochemistry and Molecular Medicine,University of Southern California

Chapters

  • 00:05Title
  • 01:09Generation of Lentivirus Particles
  • 02:34Generation of Stably Transfected Lines
  • 04:55Isolation of Monocytes from Human Blood
  • 08:00Boyden Chamber Migration Assay
  • 09:25Results: PAI-1 Promotes Macrophage Migration
  • 10:24Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol serves as a scheme for setting up a functional Tet-ON system in cancer cell lines and its subsequent use, in particular for studying the role of tumor cell-derived proteins in recruitment of monocytes/macrophages to the tumor microenvironment.

Tags

Keywords: Conditional Gene KnockdownCancer Cell LinesMonocyte RecruitmentMacrophage RecruitmentTumor MicroenvironmentLentiviral TransductionHEK-293 CellsHCT-116 CellsMDA-MB-231 CellsMonocyte PurificationPuromycin Selection

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