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Clinical & Experimental Metastasis

, Volume 25, Issue 8, pp 877–885 | Cite as

Establishment and quantitative imaging of a 3D lung organotypic model of mammary tumor outgrowth

  • Michelle D. Martin
  • Barbara Fingleton
  • Conor C. Lynch
  • Sam Wells
  • J. Oliver McIntyre
  • David W. Piston
  • Lynn M. MatrisianEmail author
Article

Abstract

The lung is the second most common site of metastatic spread in breast cancer and experimental evidence has been provided in many systems for the importance of an organ-specific microenvironment in the development of metastasis. To better understand the interaction between tumor and host cells in this important secondary site, we have developed a 3D in vitro organotypic model of breast tumor metastatic growth in the lung. In our model, cells isolated from mouse lungs are placed in a collagen sponge to serve as a scaffold and co-cultured with a green fluorescent protein-labeled polyoma virus middle T antigen (PyVT) mammary tumor cell line. Analysis of the co-culture system was performed using flow cytometry to determine the relative constitution of the co-cultures over time. This analysis determined that the cultures consisted of viable lung and breast cancer cells over a 5-day period. Confocal microscopy was then used to perform live cell imaging of the co-cultures over time. Our studies determined that host lung cells influence the ability of tumor cells to grow, as the presence of lung parenchyma positively affected the proliferation of the mammary tumor cells in culture. In summary, we have developed a novel in vitro model of breast tumor cells in a common metastatic site that can be used to study tumor/host interactions in an important microenvironment.

Keywords

Breast cancer Metastasis Microenvironment Organotypic co-culture Host–tumor interactions 

Abbreviations

GFP

Green fluorescent protein

PyVT

Polyoma virus middle T antigen

PBS

Phosphate buffered saline

H&E

Hematoxylin and eosin

FCS

Fetal calf serum

2D

Two-dimensional

3D

Three-dimensional

Notes

Acknowledgments

This work was supported by a grant from the NIH (R01-CA84360 to LMM). We are grateful to Dr. Carlos Arteaga and Dr. Shimian Qu for the pMSCV-GFP vector.

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Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Michelle D. Martin
    • 1
  • Barbara Fingleton
    • 1
  • Conor C. Lynch
    • 1
    • 2
  • Sam Wells
    • 3
  • J. Oliver McIntyre
    • 1
  • David W. Piston
    • 3
  • Lynn M. Matrisian
    • 1
    Email author
  1. 1.Department of Cancer BiologyVanderbilt UniversityNashvilleUSA
  2. 2.Department of Orthopaedics and RehabilitationVanderbilt UniversityNashvilleUSA
  3. 3.Department of Molecular Physiology and BiophysicsVanderbilt UniversityNashvilleUSA

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