A 3-D tissue model of breast morphogenesis for replacing animals in testing for endocrine disrupting substances

This project is funded by the NC3Rs which gives grants to research projects aimed at replacing, refining and reducing animals in research. Support funding was also provided by Breast Cancer UK.

MCF10A cells in 3D culture

Single MCF10A cells beginning the initial stages of collagen remodeling and structure formation. Cell actin is labeled green, the nucleus in blue and surrounding collagen in grayscale

Using human breast cells growing in artificial collagen gels we have successfully created models which can be used to watch the development of complex breast structures from single cells in real time. In addition a breast cancer model which retains hormone sensitivity in culture conditions has been used to create a sensitive test for xenoestrogenes. These chemicals can have potential endocrine disrupting effects during development and lead to an increased risk of cancer. By creating models that behave like tissue in the body a large number of animal experiments can be replaced.
Specialised microscopy techniques were used to image collagen fiber re-modelling during breast structure formation in living cells over a number of days and in some cases weeks. The video below shows a tubule forming from human MCF10A cells over a period of 5 days. Collagen fibers are coloured green. Fiber alignment takes place in the direction of cell growth and between neighbouring structures. Eventually a network of tubules is formed characteristic of breast tissue.