A novel system for mammary epithelial cell culture

A method is described for the isolation and density gradient enrichment of mammary epithelial fragments from pregnant, nonlactating bovine tissue. Immunocytochemical analysis prior to and following culture revealed specific staining with antibodies to keratin, indicating that these cells are epithelial in nature. Fragments enriched for epithelium could be stored in liquid nitrogen for extended periods prior to culture. When cast within a three-dimensional matrix of collagen gel, the mammary fragments grew as branching, duct-like structures and displayed a 4-fold increase in cell number during 10 to 12 d of culture.