Patent ID: 6828111
Filing Date: 2004-12-07
Classification: C12N

Abstract:
A method for assaying a test agent for its activity in inhibiting molecular or cellular changes characteristic of preneoplastic breast disease in a three-dimensional in vitro culture system that serves as a model for preneoplastic breast disease, said method comprising:(a) providing said three-dimensional culture system, which comprise a co-culture of (i) preneoplastic breast epithelial cells, (ii) endothelial cells and (iii) breast fibroblasts on a reconstituted basement membrane in the presence of medium containing (1) effective concentrations of growth factors that act on said epithelial and endothelial cells, and (2) effective concentrations of an estrogen such that said cells undergo morphogenesis that results in the formation of a multicellular three dimensional network of branching ductal alveolar units in culture within about 3-7 days, (b) adding said test agent to said culture system for an interval sufficient for said agent to act upon said preneoplastic breast epithelial cells, said endothelial cells or said fibroblasts (c) to a parallel control culture, adding a negative control agent for said interval, which negative control agent does not prevent or inhibit said progression, (d) examining the cultures of (b) and (c), above, for i. formation of said branching ductal alveolar units by morphology; or ii. proliferation of cells; or iii. the presence in the culture medium of one or more secreted products of said cells wherein prevention or inhibition of said ductal alveolar unit formation, said proliferation or generation of said secreted products in cultures of (a) compared to (b), is indicative that said agent has said inhibitory activity.