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Growth of seminal vesicle epithelial cells in serum-free collagen gel culture
Authors:Yasuhiro Tomooka  Stephen E Harris  John A McLachlan
Institution:(1) Development Endocrinology and Pharmacology Section, Laboratory of Reproductive and Development Toxicology, National Institute of Environmental Health Sciences, National Institutes of Health, P. O. Box 12233, 27709 Research Triangle Park, North Carolina;(2) Present address: W. Alton Jones Cell Science Center, Lake Placid, New York
Abstract:Summary Epithelial cells from mouse seminal vesicles were enzymatically dissociated enriched by gradient centrifugation, and maintained in collagen gel cultures with defined (serum-free) media. The epithelial origin of the cells was determined morhologically, immunocytochemically, and biochemically. Cells formed three-dimensional colonies with a lumen in collagen gels. Cell number was increased eight-fold within a 8 to 12-d culture period in a medium supplemented with epidermal growth factor (EGF) (10 ng/ml), insulin (10 μg/ml), transferrin (10 μg/ml), cholera toxin (10 ng/ml), and hydrocortisone (0.1 μg/ml). The cells required eGF and insulin; the growth-promoting effects of these two peptide hormones were optimized by transferrin, cholera toxin, and hydrocortisone. Fetal bovine serum did not support growth; rather, it suppressed the stimulated growth observed in serum-free media. A time-course study revealed that a lag period preceded rapi growth. The collagen gel, serum-free culture provides a powerful tool to study the effects of hormones on proliferation and differentiation of androgen sensitive cells.
Keywords:epithelial cells  seminal vesicle  serum-free media  adogen  genital tract  mouse
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