Figure 4.

Mouse mammary gland digestion and mammary epithelial cell isolation

(A) Minced mammary tissue is chopped to a sludge-like consistency to aid the digestion process.

(B) Minced mammary tissue is placed in digestion media (left panel). Following incubation, the mammary tissue digest is a translucent solution with no large tissue fragments (middle panel). Large fibrous clumps may persist in the digestion. These fibrous pieces can be manually removed using a pipette (right panel).

(C) Isolated mammary cells visualized using a light microscope. Following mammary gland digestion the sample consists of large clumps of epithelial cells and contaminating cell types such as muscle tissue and fibrous tissue (left panel, arrows). Differential centrifugation allows the purification of mammary epithelial clumps (middle panel). The epithelial clumps can be further digested to single cells for FACS (right panel). Scale bar, 0.25 mm.