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Combined Conditional Knockdown and Adapted Sphere Formation Assay to Study a Stemness-Associated Gene of Patient-derived Gastric Cancer Stem Cells
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JoVE Journal Pesquisa do Câncer
Combined Conditional Knockdown and Adapted Sphere Formation Assay to Study a Stemness-Associated Gene of Patient-derived Gastric Cancer Stem Cells

Combined Conditional Knockdown and Adapted Sphere Formation Assay to Study a Stemness-Associated Gene of Patient-derived Gastric Cancer Stem Cells

DOI:
10.3791/60799-v

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09:24 min

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May 09, 2020

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, , ,
1Guangdong Key Laboratory of Genome Stability and Human Disease Prevention, Department of Pharmacology and Shenzhen University International Cancer Center,Shenzhen University School of Medicine

Capítulos

  • 00:04Introduction
  • 01:02Generation of Inducible Knockdown GCSCs Lines
  • 03:31Generation of Stable Transfected Cell Lines
  • 06:12Sphere Formation Assay
  • 06:54Results: Inducible Knockdown of Clusterin Inhibits GCSC Self-renewal Capacity
  • 08:22Conclusion

Summary

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Tadução automática

In this protocol, we present an experimental design using a conditional knockdown system and an adapted sphere formation assay to study the effect of clusterin on the stemness of patient-derived GCSCs. The protocol can be easily adapted to study both in vitro and in vivo function of stemness-associated genes in different types of CSCs.

Tags

Cancer Stem CellsConditional KnockdownSphere Formation AssayStemness-associated GenesGastric CancerLentivirus293T CellsTransfectionVirus Concentration

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