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

10:26 min

October 21, 2022

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Kapitel

  • 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

Automatische Übersetzung

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.

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