* *PBS (sterile)
- Remove mammary glands from mice and place in sterile PBS to rinse.
- Chop/mince glands with razor blade and place into 15ml conical with 10mls collagenase solution. (Typically 4-8 virgin glands/tube ~500-1000mg tissue/tube)
- Digest tissue in collagenase solution for 2-3 hours at 37oC on a rotator.
- Transfer supernatant (this contains single cells/stromal cells) to a new tube and leave the settled material in the tube (epithelial cells). Wash the remaining settled material by adding 10mls of wash solution. Spin cells.
- Repeat the wash 3X.
The primary cells are now ready to expand in tissue culture in appropriate media.