title: Culture of bovine ovarian follicle wall sections maintained the highly estrogenic profile under basal and chemically defined conditions.
abstract: Follicle cultures reproduce in vitro the functional features observed in vivo. In a search for an ideal model, we cultured bovine antral follicle wall sections (FWS) in a serum-free defined medium (DM) known to induce 17β-estradiol (E2) production, and in a nondefined medium (NDM) containing serum. Follicles were sectioned and cultured in NDM or DM for 24 or 48 h. Morphological features were determined by light microscopy. Gene expression of steroidogenic enzymes and follicle-stimulating hormone (FSH) receptor were determined by RT-PCR; progesterone (P4) and E2 concentrations in the media were measured by radioimmunoassay. DM, but not NDM, maintained an FWS morphology in vitro that was similar to fresh tissue. DM also induced an increase in the expression of all steroidogenic enzymes, except FSH receptor, but NDM did not. In both DM and NDM, there was a gradual increase in P4 throughout the culture period; however, P4 concentration was significantly higher in NDM. In both media, E2 concentration was increased at 24 h, followed by a decrease at 48 h. The E2:P4 ratio was higher in DM than in NDM. These results suggest that DM maintains morphological structure, upregulates the expression of steroidogenic enzyme genes, and maintains steroid production with a high E2:P4 ratio in FWS cultures.
authors: Universidade de Brasília, Instituto de Ciências Biológicas, Laboratório de Biotecnologia da Reprodução, Departamento de Ciências Fisiológicas, BrasíliaDF, Brasil.
authors: R B
authors: RB
authors: Vasconcelos
authors: L P
authors: LP
authors: Salles
authors: I
authors: I
authors: Oliveira e Silva
authors: L V M
authors: LV
authors: Gulart
authors: D K
authors: DK
authors: Souza
authors: F A G
authors: FA
authors: Torres
authors: A L
authors: AL
authors: Bocca
authors: A A M
authors: AA
authors: Rosa e Silva
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D001141:Aromatase / Q000235:genetics; D002417:Cattle; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000235:genetics; D003470:Culture Media / Q000494:pharmacology*; D016895:Culture Media, Serum-Free; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D015870:Gene Expression; D006080:Ovarian Follicle / Q000033:anatomy & histology / Q000187:drug effects*; D010750:Phosphoproteins / Q000235:genetics; D011374:Progesterone / Q000494:pharmacology*; D011375:Progesterone Reductase / Q000235:genetics; D011962:Receptors, FSH / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013254:Steroid 17-alpha-Hydroxylase / Q000235:genetics; D046509:Tissue Culture Techniques; D000098682:Steroidogenic Acute Regulatory Protein
chemical_list: D003470:Culture Media; D016895:Culture Media, Serum-Free; D010750:Phosphoproteins; D011962:Receptors, FSH; D000098682:Steroidogenic Acute Regulatory Protein; D011374:Progesterone; D004958:Estradiol; D011375:Progesterone Reductase; D001141:Aromatase; D013254:Steroid 17-alpha-Hydroxylase; D002786:Cholesterol Side-Chain Cleavage Enzyme
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