Development of three-dimensional culture models to study cancer development and metastasis

Bray LJ1, Murekatete B1, Jaeschke A1, Koch M1, Tsurkan MV2, Risbridger G3, Werner C2 and Hutmacher DW1

  1. Centre for Regenerative Medicine, Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Queensland, Australia.
  2. Leibniz Institute for Polymer Research, Max Bergmann Center of Biomaterials, Dresden, Saxony, Germany.
  3. Monash University, Clayton, Victoria, Australia.

The culture of cells on rigid 2D substrates, such as tissue culture plastic, does not recreate the dynamic and highly complex tissue microenvironment, but rather distorts cell-integrin and cell-cell interactions, affecting gene expression, signal transduction, cell proliferation and differentiation, and thus is physiologically irrelevant. Naturally, cells are embedded in an extracellular matrix (ECM) that provides not only architectural support, but also chemical and mechanical signals to cells in vivo. Hydrogels prepared from star-shaped poly(ethylene glycol) (PEG) and maleimide-functionalised heparin provide a potential matrix for use in developing three dimensional (3D) models. We have previously demonstrated that these hydrogels support tri-cultures of human umbilical vein endothelial cells (HUVECs) with mesenchymal stromal cells (MSCs) and breast or prostate cancer epithelial cell lines. We extended this body of work to study the effects of cancer associated fibroblasts (CAFs) on tumour angiogenesis. Also, we investigated the ability to produce a tri-culture mimicking tumour angiogenesis with epithelial cells, and tissue-specific microvascular endothelial cells and fibroblasts. Cultures were analysed via immunostaining and observed using confocal microscopy. We demonstrated the ability of starPEG-heparin hydrogels to support co- and tri-cultures of primary patient-derived prostate or breast cells, with capillary formation by the endothelial cells. Interactions were visualised between all cell types via confocal microscopy. Our results confirm the suitability of hydrogels constructed from starPEG-heparin for the co-cultivation of primary patient-derived tissue specific cells to study cell-cell and cell-matrix interactions in a 3D microenvironment. This represents a step forward in the development of 3D culture models to study the pathomechanisms of various cancer types.