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Related Experiment Video

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Rat Mesentery Angiogenesis Assay
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Implementation of a basement membrane invasion assay using mesenteric tissue.

Ritobrata Ghose1, Alistair J Rice1, Ernesto Cortes1

  • 1Cellular and Molecular Biomechanics Laboratory, Department of Bioengineering, Imperial College London, London, United Kingdom.

Methods in Cell Biology
|April 27, 2020
PubMed
Summary

This study introduces a novel 3D in vitro assay using mouse mesentery to accurately model basement membrane invasion, a key step in cancer metastasis. The assay and its quantification tool, Q-Pi, offer a more physiologically relevant method for studying tumor cell invasion.

Keywords:
3D reconstructionBasement membraneCell invasionInvasion assayMesenteryMetastasisQuantification of invasion

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Area of Science:

  • Oncology
  • Cell Biology
  • Biomaterials

Background:

  • Metastasis, responsible for 90% of cancer deaths, critically involves epithelial tumor cells invading the basement membrane (BM).
  • Existing in vitro assays often lack the physiological accuracy needed to study BM invasion effectively.
  • Developing accurate models is crucial for understanding and combating cancer spread.

Purpose of the Study:

  • To introduce a novel 3D in vitro assay that accurately mimics the physiological basement membrane (BM) for studying cancer cell invasion.
  • To validate the use of mouse mesenteric tissue as a physiologically relevant BM model.
  • To present a user-friendly tool for quantifying cellular invasion within this 3D model.

Main Methods:

  • Development of a cost-effective protocol for extracting and preparing mouse mesenteric tissue as a basement membrane (BM) mimic.
  • Characterization of the mesenteric tissue's key BM components: type IV collagen, laminin-1, and perlecan.
  • Introduction of Q-Pi, a software tool for 3D reconstruction, visualization, and quantification of cellular invasion.

Main Results:

  • The mouse mesenteric tissue accurately recapitulates the essential components of the epithelial basement membrane (BM).
  • The developed 3D in vitro assay provides a physiologically relevant platform for studying tumor cell invasion.
  • The Q-Pi tool enables efficient and detailed cellular-level quantification of invasion.

Conclusions:

  • The novel 3D invasion assay using mouse mesentery offers a physiologically accurate model for studying basement membrane (BM) invasion in cancer.
  • This assay overcomes limitations of previous in vitro methods, providing a more realistic simulation of tumor cell metastasis.
  • The integrated Q-Pi tool enhances the study of cellular invasion dynamics, aiding cancer research.