Development of 3D culture reagents and scoring systems for assays of normal andmalignant epithelial stem cells
The overarching objective of this proposal is to develop and validate a new, robust and reproducible 3D culture system that supports the clonal growth of primary sources of normal and malignant human epithelial cells and that is suitable for commercialization by STEMCELL Technologies who will partner in its development. The work will systematically address 3 Specific Aims. These will be designed to test, optimize and validate the culture components required to generate clonal structures that mimic in vivo outputs of freshly isolated human epithelial cells at high efficiency, reproducibly, and with a described scoring method that may be adapted to automation for future use for screening molecules that promote or inhibit growth. This project will use cells isolated from normal human mammary tissue and samples of human breast cancer as a prototype, but we expect that the general approach will be transferable to other types of epithelial cells with minimal modification. Specific Aims: 1. To develop and characterize an initial basic system for growing defined subsets of normal human mammary epithelial progenitors in 3D Matrigel cultures.