title: Purification of Distinct Subsets of Epithelial Cells from Normal Human Breast Tissue.
abstract: The mammary epithelium is composed of a variety of specialized cell types that function in a coordinated fashion to produce and eject milk through multiple cycles of pregnancy. The ability to identify and purify these subsets of cells in order to interrogate their growth and differentiation capacities, as well as to characterize the molecular mechanisms that regulate their behavior, is essential in identifying the processes associated with breast cancer initiation and progression. This methods chapter outlines the step-by-step methods for dissociating human breast reduction specimens to a single cell suspension of viable cells. As well, strategies for purifying four distinct subsets of epithelial cells by using fluorescence-activated cell sorting and protocols for interrogating the growth and differentiation properties of these purified cells at clonal densities in adherent culture are also described.
authors: Cancer Research UK Cambridge Institute, Li Ka Shing Centre, University of Cambridge, Robinson Way, Cambridge, CB2 0RE, UK. mona.shehata@uhnresearch.ca.
authors: Mona
authors: M
authors: Shehata
authors: Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge, UK.|STEMCELL Technologies Inc, Vancouver, British Columbia, Canada.
authors: John
authors: J
authors: Stingl
mesh_terms: D000818:Animals; D001940:Breast / Q000166:cytology*; D001943:Breast Neoplasms / Q000473:pathology; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology*; D004848:Epithelium / Q000502:physiology; D005260:Female; D006801:Humans; D042361:Mammary Glands, Human / Q000166:cytology*; D051379:Mice; D041681:NIH 3T3 Cells
keywords: Cell culture; Colony-forming cells; Flow cytometry; Human breast tissue; Stem cells