In vitro multipotent differentiation and barrier function of a human mammary epithelium.pdf

REGULAR ARTICLE In vitro multipotent differentiation and barrier function of a human mammary epithelium Aaron M. Marshall Vaibhav P. Pai Maureen A. Sartor Nelson D. Horseman Received: 2 April 2008 /Accepted: 6 October 2008 / Published online: 13 November 2008 # Springer-Verlag 2008 Abstract As demonstrated by a variety of animal studies, barrier function in the mammary epithelium is essential for a fully functioning and differentiated gland. However, there is a paucity of information on barrier function in human mammary epithelium. Here, we have established character- istics of a polarizing differentiating model of human mammary epithelial cells capable of forming a high- resistance/low-conductance barrier in a predictable manner, viz., by using MCF10A cells on permeable membranes. Inulin flux decreased and transepithelial electrical resistance (TEER) increased over the course of several days after seeding MCF10A cells on permeable membranes. MCF10A cells exhibited multipotent phenotypic differenti- ation into layers expressing basal and lumenal markers when placed on permeable membranes, with at least two distinct cell phenotypes. A clonal subline of MCF10A, generated by culturing stem-like cells under non-adherent conditions, also generated a barrier-forming epithelial membrane with cells expressing markers of both basal and lumenal differentiation (CD10 and MUC1, respectively). Progressive changes associated with differentiation, includ- ing wholesale inhibition of cell-cycle genes and stimulation of cell and tissue morphogenic genes, were observed by gene expression profiling. Clustering and gene ontology categorization of significantly altered genes revealed a pattern of lumenal epithelial-cell-specific differentiation. Keywords Breast . Transepithelial electrical resistance . Tight junctions .Milk protein .MCF10A . Gene expression profile . Human Introduction Postnatal mammary gland development is controlled by interactions between endocrine hormones and fa