epidermal-growth-factor and phosphorylethanolamine

When human epithelial cells that had grown out from either carcinoma or histologically non-malignant breast tissues were seeded within type I collagen gels in serum-free medium, they successively grew and protruded many radial duct-like extensions with lumina. Separate deletion of each of the supplements from the medium showed that growth as well as morphological differentiation of carcinoma-derived cells were prevented in the absence of epidermal growth factor (EGF) or hydrocortisone. Removal of insulin or ethanolamine plus phosphoethanolamine caused a significant inhibition of cell growth without interfering with the morphological differentiation. Contrary to the case with carcinoma-derived cells, both growth and morphological differentiation of epithelial cells derived from non-malignant breast tissues were prevented when EGF, hydrocortisone or insulin was absent. Removal of each of the other supplements (except for transferrin), including ethanolamine plus phosphoethanolamine, prolactin, or prostaglandin, caused a significant inhibition of cell growth with no apparent inhibition of morphological differentiation. The present results suggest that human epithelial cells derived from either carcinoma or histologically non-malignant breast tissues strongly depend on the presence of EGF and hydrocortisone and there is a decreased dependence on insulin in carcinoma-derived cells with respect to their growth and morphological differentiation during culture within collagen gels.

Topics: Alprostadil; Breast Neoplasms; Cell Differentiation; Cell Division; Collagen; Culture Media; Epidermal Growth Factor; Epithelium; Ethanolamine; Ethanolamines; Female; Humans; Hydrocortisone; Insulin; Prolactin; Transferrin; Tumor Cells, Cultured

Extracts of term human placenta were tested for enhancement of proliferative growth of primary cultures of human keratinocytes. Saline extracts or supernatants from homogenates were dialyzed extensively, lyophilized, and tested in subcultures of keratinocytes in MCDB 153 medium with 0.1 mM Ca++ containing only defined supplements (insulin, hydrocortisone, transferrin, ethanolamine, phosphoethanolamine). Cells plated in the absence of EGF at moderately high densities (1000-3000 cells per cm2) formed colonies and grew in the presence of placental extract at 25-500 micrograms/ml. Extracts of cord serum or maternal serum were inactive, suggesting that the activity is derived from placental tissue. The activity is not EGF, since the activity in the placental extract, unlike EGF, did not promote growth at low cell density, was synergistic with EGF under some conditions, and did not produce changes in colonial morphology which occurred in the presence of EGF. Unlike keratinocyte growth-promoting activity in bovine hypothalamic extract, the activity is non-dialyzable and is destroyed at 100 degrees C. Placental extract could not replace any of the defined components of the medium and is therefore distinct from them. The presence of activity in the placenta with distinctive properties suggests that this is a previously undescribed material with growth-promoting properties for epithelium.

Topics: Cell Count; Cell Division; Culture Media; Dialysis; Epidermal Cells; Epidermal Growth Factor; Ethanolamine; Ethanolamines; Female; Fetal Blood; Growth Substances; Hot Temperature; Humans; Hydrocortisone; Insulin; Keratins; Male; Placenta; Pregnancy; Transferrin

A number of single-cell-cloned cell lines have been used to examine the growth-promoting effects of putative mammotrophic agents on the various cell types in normal and neoplastic rat mammary glands. A partially purified novel pituitary-derived growth factor stimulates only cuboidal epithelial cells to divide whereas fibroblast growth factor (FGF) stimulates the growth of stromal and myoepithelial-like cells. Epidermal growth factor (EGF) has a widespread but variable growth-stimulating action, but prolactin and growth hormone are essentially inactive when added alone at a concentration of 5 micrograms/ml. Phosphoethanolamine stimulates the growth of one epithelial cell line and a derivative myoepithelial-like cell line, but is inactive on the other cell lines tested. The use of defined cloned cell lines provides a direct and reproducible assay for the identification and purification of inducers of mammary growth.

Topics: Animals; Cell Division; Cells, Cultured; Epidermal Growth Factor; Epithelial Cells; Ethanolamines; Fibroblast Growth Factors; Growth Substances; Mammary Glands, Animal; Pituitary Hormones; Rats