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

• Published in: Cell and tissue research 2009-02, Vol.335 (2), p.383-395
• Main Authors: Marshall, Aaron M, Pai, Vaibhav P, Sartor, Maureen A, Horseman, Nelson D
• Format: Article
• Language: English
• Subjects: Adherens Junctions - physiology; Biomedical and Life Sciences; Biomedicine; Breast; Cadherins - metabolism; Cell culture; Cell cycle; Cell Differentiation - genetics; Cell Differentiation - physiology; Cell Line; Cell Polarity - drug effects; Cell Polarity - physiology; Cellular biology; dairy protein; Down-Regulation - genetics; Electric Impedance; Epithelial Cells - cytology; Epithelial Cells - drug effects; Epithelial Cells - physiology; Gene expression profile; Gene Expression Profiling; Genotype & phenotype; Human Genetics; Humans; Mammary Glands, Human - cytology; Mammary Glands, Human - drug effects; Mammary Glands, Human - physiology; MCF10A; Membrane Proteins - metabolism; Models, Biological; Molecular Medicine; Mucin-1 - metabolism; Neprilysin - metabolism; Occludin; Proteomics; Regular Article; tight junctions; Tight Junctions - physiology; Transepithelial electrical resistance; Tretinoin - pharmacology; Up-Regulation - genetics
• ISSN: 0302-766X; 1432-0878
• DOI: 10.1007/s00441-008-0719-0

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 characteristics 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 differentiation 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, including 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.