Ovary culture of Goldfish Carassius auratus

In this study primary culture, subculture1 and subculture2 of ovarian tissue from goldfish was done to develop an in vitro system of ovarian cells in this species. A goldfish weighing 42g and 7 cm in length was anesthetized by Clove powder (0.2 g/lit) and its ovary was removed and put in a petri dish containing 5ml L-15 medium, Streptomycin sulphate (200 ug/ml), Penicillin G potassium (Gibco, 200 iu/ml) and Amphotericin B (5 ug/ml). The oocytes which were in advanced maturation stage (IV) were isolated from ovary and the ovary cut into small pieces (explants) by a 5ml syringe. The ovary explants cultivated in 5ml L-15 medium supplemented with 15% FBS, Streptomycin sulphate (100 ug/ml), Penicillin G potassium (Gibco, 100 iu/ml) and Amphotericin B (2.5 ug/ml) at 25°C. After two weeks when confluent monolayer formed on the bottom of the flask, the cells were washed with PBS containing antibiotics, dislodged by Trypsin-EDTA solution (0.25%) and cultured in 5ml of L-15 medium, FBS (15%), antibiotics at 25 °C (first subculture). After confluent monolayer formation, on the sixteenth day similar procedure was followed for the second subculture.The emerging cells from explants and two subcultures exhibited fibroblast-like cells.