An Ex vivo Model to Study Hormone Action in the Human Breast
The study of hormone action in the human breast has been hampered by lack of adequate model systems. Upon in vitro culture, primary mammary epithelial cells tend to lose hormone receptor expression. Widely used hormone receptor positive breast cancer cell lines are of limited relevance to the in vivo situation. Here, we describe an ex vivo model to study hormone action in the human breast. Fresh human breast tissue specimens from surgical discard material such as reduction mammoplasties or mammectomies are mechanically and enzymatically digested to obtain tissue fragments containing ducts and lobules and multiple stromal cell types. These tissue microstructures kept in basal medium without growth factors preserve their intercellular contacts, the tissue architecture, and remain hormone responsive for several days. They are readily processed for RNA and protein extraction, histological analysis or stored in freezing medium. Fluorescence activated cell sorting (FACS) can be used to enrich for specific cell populations. This protocol provides a straightforward, standard approach for translational studies with highly complex, varied human specimens.
Keywords: Medicine ; Issue 95 ; Hormone signaling ; breast cancer ; reduction mammoplasty ; breast tissue microstructures ; ex vivo model ; estrogen ; progesterone ; mammary epithelial cells ; tissue digestion ; paracrine signaling ; microenvironment ; tissue architecture
Record created on 2015-02-09, modified on 2016-08-09