Growth of seminal vesicle epithelial cells in serum-free collagen gel culture.
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 morphologically, 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 micrograms/ml), transferrin (10 micrograms/ml), cholera toxin (10 ng/ml), and hydrocortisone (0.1 microgram/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 rapid 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.
D000818:Animals; D001790:Blood Physiological Phenomena; D002455:Cell Division / Q000187:drug effects; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology; D003470:Culture Media / Q000032:analysis / Q000494:pharmacology*; D004847:Epithelial Cells; D005782:Gels; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D012669:Seminal Vesicles / Q000166:cytology*
D003470:Culture Media; D005782:Gels; D003094:Collagen