A heterogeneous in vitro three dimensional model of tumour-stroma interactions regulating sprouting angiogenesis.

PloS One
Pedro Correa de SampaioWilliam R English

Abstract

Angiogenesis, the formation of new blood vessels, is an essential process for tumour progression and is an area of significant therapeutic interest. Different in vitro systems and more complex in vivo systems have been described for the study of tumour angiogenesis. However, there are few human 3D in vitro systems described to date which mimic the cellular heterogeneity and complexity of angiogenesis within the tumour microenvironment. In this study we describe the Minitumour model--a 3 dimensional human spheroid-based system consisting of endothelial cells and fibroblasts in co-culture with the breast cancer cell line MDA-MB-231, for the study of tumour angiogenesis in vitro. After implantation in collagen-I gels, Minitumour spheroids form quantifiable endothelial capillary-like structures. The endothelial cell pre-capillary sprouts are supported by the fibroblasts, which act as mural cells, and their growth is increased by the presence of cancer cells. Characterisation of the Minitumour model using small molecule inhibitors and inhibitory antibodies show that endothelial sprout formation is dependent on growth factors and cytokines known to be important for tumour angiogenesis. The model also shows a response to anti-angiogen...Continue Reading

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Methods Mentioned

BETA
FCS
protein assay
transmission electron microscopy
confocal microscopy
electron microscopy
dissect

Software Mentioned

GraphPad Prim
LAS AF Leica
Metamorph
Volocity®
Living Image®
GraphPad
Volocity
Deben
ImageJ

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