ABSTRACT
Normal human breast lobules, freshly isolated by precision microdissection of tissue stained with methylene blue chloride, were assayed for their ability to induce neovascularization (angiogenesis) in rabbit irises. Histologically, normal lobules from cancerous breast induced angiogenesis twice as often as lobules from noncancerous breasts, suggesting that preneoplastic transformation is diffuse.
Subject(s)
Breast/physiology , Iris/blood supply , Neovascularization, Pathologic , Precancerous Conditions/physiopathology , Age Factors , Animals , Female , Humans , Middle Aged , Rabbits , Time FactorsABSTRACT
Human mammary epithelial cells were dissociatd from mastectomy tissues. The contaminating fibroblasts were removed by the use of Percoll density-gradient centrifugation, which utilizes the difference in buoyant densities between epithelial cells and fibroblasts. A preparation highly enriched for mammary epithelial cells ws then embedded in collagen gel and cultured in Ham's F12 medium containing 12.5% horse serum, 2.5% fetal calf serum, 0.1 microgram cholera toxin/ml, an extract prepared from human male urine (L microgram protein/ml), and a hormone combination of 10 microgram insulin/ml, 10 microgram human placental lactogen/ml, 1 microgram aldosterone/ml, and 0.5 microgram hydrocortisone/ml. Sustained growth leading to an increase of tenfold to thirtyfold in cell number over the initial value was accomplished in primary culture, and this growth was maintained even after passage to secondary culture. Deletion of either the urine extract or the hormone combination resulted in less than optimal growth. Subsequent studies showed that hydrocortisone alone could replace the hormone combination. In addition, urine extract could be replaced by extracts prepared from human kidneys or brains. The collagen gel system provies a reproducible and consistent method for sustained three-dimensional growth of mammary epithelial cells from human breast tissue in primary as well as passaged cultures.
Subject(s)
Breast/cytology , Cells, Cultured , Gels , Aldosterone , Cholera Toxin , Collagen , Culture Media , Epithelial Cells , Estradiol , Humans , Hydrocortisone , Insulin , ProgesteroneABSTRACT
Persistent human mammary lobules (PL) remaining after the menopause, and certain atypical lobules (ALA) are morphologically similar to the common preneoplastic hyperplastic alveolar nodule (HAN) of mice of strains having a high incidence of mammary carcinoma. This and other evidence suggest that like the HAN of mice human PL and ALA are precancerous.
