Isolation of the ovarian follicular cells in common carp and evaluation of their endocrine activity in primary cell culture

Objective: To study endocrine activity of isolated carp ovarian follicular cells (granulose and theca cells). Progesterone (P4) and estrogen (E2) levels in the culture media of cultivated carp ovarian follicular cells were estimated using radioimmunoassay (RIA). Method & Materials: Oocytes were isolated from the ovaries of female carps. Interstitial tissue was mechanically removed in order to obtain single oocytes surrounded only by the follicular envelope. Such a preparation was trypsinized at room temperature and also with greater success at 37◦C. The cells were grown as monolayer in 24- well microplates in MEM medium supplemented with calf serum. Culture media were analyzed for estrogen and progesterone content by appropriate radioimmunoassay. Results & Conclusion: Trypsinization of ovarian follicles resulted in the formation of the cell suspension contained both cells and cell clumps. It was the mixture of theca (T) and granulose (G) cells. The follicular cells attached to glass and grew well during culture period. The temperature of 20◦C was preferable for the successful growth of the cells in tissue culture. E2 was the main steroid hormone secreted by cultivated cells.