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Assessment of Mitochondrial Health in Cancer-Associated Fibroblasts Isolated from 3D Multicellular Lung Tumor Spheroids
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JoVE Journal Cancer Research
Assessment of Mitochondrial Health in Cancer-Associated Fibroblasts Isolated from 3D Multicellular Lung Tumor Spheroids

Assessment of Mitochondrial Health in Cancer-Associated Fibroblasts Isolated from 3D Multicellular Lung Tumor Spheroids

DOI:
10.3791/64315-v

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10:26 min

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October 21, 2022

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, , ,
1Department of Biomedical Engineering,Indian Institute of Technology Ropar

Chapters

  • 00:04Introduction
  • 01:13Preparation of Cell Suspension
  • 03:24Live-Dead Analysis of Tumor Spheroids
  • 04:05Disintegration and Cell Suspension of Tumor Spheroids
  • 05:02Cancer-Associated Fibroblast (CAF) Isolation Through Microbeads
  • 06:19Flow Cytometry-Based Analysis of ACTA2 Expression in Isolated CAFs
  • 07:28Results: Assessment of Mitochondrial Health in Cancer-Associated Fibroblasts
  • 09:30Conclusion

Summary

Automatic Translation

Automatic Translation

Multicellular 3D tumor spheroids were prepared with lung adenocarcinoma cells, fibroblasts, and monocytes, followed by the isolation of cancer-associated fibroblasts (CAFs) from these spheroids. Isolated CAFs were compared with normal fibroblasts to assess mitochondrial health by studying the mitochondrial transmembrane potential, reactive oxygen species, and enzymatic activities.

Tags

Keywords: Cancer-associated FibroblastsCAF3D Multicellular Lung Tumor SpheroidsMitochondrial HealthTumor MicroenvironmentIn Vitro ModelCell IsolationCell CultureA549 CellsMRC-5 CellsTHP-1 CellsTumor Spheroid Formation

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