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 media were collected after 72 h and frozen at -20°C. Carp ovarian follicles also were separated mechanically and incubated in BSS [basic salt solution] Cortland medium in 24-well microplates for 48 h at 20°C. Then, they were divided into two groups control group which were incubated in BSS medium and experimental group which subdivided into three subgroups according to treatment with different concentration of collected pituitary secretion [50, 100 and 200 micro 1/ml]. Follicles culture media were collected 24 h later and were analyzed for 17-beta-oestradiol [E[2]] and 17-alpha-hydroxy progesterone [P[4]] content by radioimmunoassay [RIA]. The results showed that adding low concentration [50 micro 1/ml] of collected pituitary secretion [CPS] increased steroid hormones [E[2] and P[4]] secretion of incubated ovarian follicles significantly [P<0.05] but the high concentration of CPS [200 micro 1/ml] significantly decreased the secretion of E[2] and P[4] [P<0.05]. Collected pituitary secretion at the concentration of 100 micro 1/ml had no significant effect on steroid hormones [P>0.05]