Physiologic characterization of transformed and cloned rat granulosa cells.
Properties of a clonal line of SV40-transformed rat granulosa cells (DC3 cells) were elucidated. DC3 cells were maintained in vitro in Iscove Modified Dulbecco Medium that contained 20% fetal bovine serum. The cells had a logarithmic growth phase doubling time of approximately 18 h and produced detectable quantities of estrone, estradiol, and progesterone. Steroidogenesis was increased by supplementation with either steroidogenic substrates or agents that stimulated activity of adenylate cyclase. Production of progesterone and estrogens was enhanced when medium was supplemented with 25-hydroxycholesterol, and production of estradiol was enhanced by medium supplementation with androstenedione. Treatments with forskolin and cholera toxin resulted in marked increases of cyclic adenosine 3',5'-monophosphate (cAMP) in medium and cells and enhanced steroidogenesis. Isoproterenol and vasoactive intestinal peptide, but not follicle-stimulating hormone (FSH), luteinizing hormone (LH), insulin or prolactin, stimulated cAMP secretion by suspended cells. DC3 cells had small but detectable levels of binding to FSH, but binding of LH and epidermal growth factor could not be detected. DC3 cells possess characteristics expected of granulosa cells arrested in an early stage of differentiation and may provide a useful model for studies of "immature" granulosa cell functions.
Fitz
T A
TA
Department of Obstetrics and Gynecology, Uniformed Services, University of the Health Sciences, Bethesda, Maryland 20814.
Wah
R M
RM
Schmidt
W A
WA
Winkel
C A
CA
D000818:Animals; D002454:Cell Differentiation; D002461:Cell Line, Transformed; D002999:Clone Cells; D000242:Cyclic AMP / Q000096:biosynthesis; D005260:Female; D005640:Follicle Stimulating Hormone / Q000378:metabolism; D006107:Granulosa Cells / Q000032:analysis / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D006888:Hydroxycholesterols / Q000494:pharmacology; D066298:In Vitro Techniques; D007545:Isoproterenol / Q000494:pharmacology; D007986:Luteinizing Hormone / Q000378:metabolism; D008954:Models, Biological; D011863:Radioimmunoassay; D051381:Rats; D011956:Receptors, Cell Surface / Q000032:analysis; D013256:Steroids / Q000096:biosynthesis
D006888:Hydroxycholesterols; D011956:Receptors, Cell Surface; D013256:Steroids; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone; D000242:Cyclic AMP; D007545:Isoproterenol