A three-dimensional co-culture system to investigate macrophage-dependent tumor cell invasion


  • Amy R. Dwyer School of Medicine and Pharmacology, University of Western Australia
  • Lesley G. Ellies School of Anatomy, Physiology and Human Biology, University of Western Australia
  • Andrea L. Holme Centre for Microscopy, Characterisation and Analysis, University of Western Australia
  • Fiona J. Pixley School of Medicine and Pharmacology, University of Western Australia




macrophage, motility, 3D co-culture, invasion, mammosphere


Macrophages infiltrate cancers and promote progression to invasion and metastasis. To directly examine tumor-associated macrophages (TAMs) and tumor cells interacting and co-migrating in a three-dimensional (3D) environment, we have developed a co-culture model that uses a PyVmT mouse mammary tumor-derived cell line and mouse bone marrow-derived macrophages (BMM). The Py8119 cell line was cloned from a spontaneous mammary tumor in a Tg(MMTV:LTR-PyVmT) C57Bl/6 mouse and these cells form 3-dimensional (3D) spheroids under conditions of low adhesion. Co-cultured BMM infiltrate the Py8119 mammospheres and embedding of the infiltrated mammospheres in Matrigel leads to subsequent invasion of both cell types into the surrounding matrix. This physiologically relevant co-culture model enables examination of two critical steps in the promotion of invasion and metastasis by BMM: 1) macrophage infiltration into the mammosphere and, 2) subsequent invasion of macrophages and tumor cells into the matrix. Our methodology allows for quantification of BMM infiltration rates into Py8119 mammospheres and demonstrates that subsequent tumor cell invasion is dependent upon the presence of infiltrated macrophages. This method is also effective for screening macrophage motility inhibitors. Thus, we have developed a robust 3D in vitro co-culture assay that demonstrates a central role for macrophage motility in the promotion of tumor cell invasion.


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How to Cite

Dwyer AR, Ellies LG, Holme AL, Pixley FJ. A three-dimensional co-culture system to investigate macrophage-dependent tumor cell invasion. J Biol Methods [Internet]. 2016Jul.24 [cited 2021May8];3(3):e49. Available from: https://jbmethods.org/jbm/article/view/132




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