Media, Enzymes, & Supplements
The human mammary gland is a compound tubulo-alveolar gland composed of a series of branched ducts that drain sac-like alveoli. These ducts and alveoli are composed of two general lineages of epithelial cells: the luminal cells and the underlying smooth muscle-like myoepithelial cells. Recent reports have demonstrated the presence of the phenotypically distinct progenitor and non-progenitor epithelial cell subpopulations within the mammary epithelium.1-5 Subsets of progenitors include the bipotent progenitor (EpCAM+MUC1-CD49f+CD10+) which can generate both luminal and myoepithlial cells, and the luminal-restricted progenitor (EpCAM+MUC1+CD49f+CD10-). These distinct subpopulations can be isolated from normal mammary tissue or from mammary epithelial cell cultures using either flow cytometry or immunomagnetic cell separation. The growth and differentiation potential of these cells can be monitored by their ability to form colonies when plated at low density in vitro.
mammary epithelial cell enrichment and culture
StemCell Technologies offers products for dissociation of human and mouse mammary tissue, the enrichment of subsets of human mammary epithelial cell progenitors, and the culture and characterization of human and mouse mammary epithelial cells. Single cell suspensions of mammary cells can be obtained by a two-step process:
The tissue is enzymatically dissociated into mammary organoids with a collagenase/hyaluronidase cocktail.
The organoids are dissociated into a single cell suspension using trypsin, DNase, and dispase.
Purified subsets of human cells can be isolated from the single cell suspension by flow cytometry or by immunomagnetic separation using EasySep?/SUP> based on the differential expression of epithelial cell adhesion molecule (EpCAM), CD49f, CD10 and the MUC1 glycoprotein. Purified mouse mammary epithelial cells can be isolated by using EasySep?/SUP> to remove the non-epithelial cells, followed by FACS to enrich for cells expressing CD24 and/or CD49f.
Both purified and non-purified mammary epithelial cells can be cultured on tissue culture plastic or within 3-dimensional collagen gels using EpiCult?B, a specially formulated medium designed to support the proliferation of human and mouse mammary epithelial cells.