Pituitary primary cell culture of Common carp (Cyprinus Carpio) and evaluation of its secretion effect on endocrine activity of incubated ovarian follicles

Abstract

In this study, five carp pituitary glands were collected and dispersed enzymatically and mechanically.Then, the cells were cultivated as monolayer in MEM (minimum essential medium Eagle). The culture mediawere collected after 72 h and frozen at -20°C. Carp ovarian follicles also were separated mechanically andincubated in BSS (basic salt solution) Cortland medium in 24-well microplates for 48 h at 20°C. Then, theywere divided into two groups: control group which were incubated in BSS medium and experimental groupwhich subdivided into three subgroups according to treatment with different concentration of collectedpituitary secretion (50, 100 and 200 μl/ml). Follicles culture media were collected 24 h later and wereanalyzed for 17-β-oestradiol (E2) and 17-α-hydroxy progesterone (P4) content by radioimmunoassay (RIA).The results showed that adding low concentration (50 μl/ml) of collected pituitary secretion (CPS) increasedsteroid hormones (E2 and P4) secretion of incubated ovarian follicles significantly (P