In the present study, a piece of the ovary of a 6 years old female Sterlet sturgeon (A. ruthenus), a farmed fish (729 g and 47.3 cm), was removed after anaesthetizing with clove carpel powder (0.4 g/lit). Ovarian follicles were isolated from each other and were washed using sterile phosphate-buffered saline (PBS) containing antibiotics. Follicular cells were then separated by treating oocytes with 0.25% Trypsin-EDTA in Ca2+ and Mg2+ free PBS and were cultured in medium L-15 supplemented with 20% FBS at 22°C (1.25×106cells/ml). After twelve days, when confluent monolayer cells covered the bottom surface of the flask, the cells were dislodged by Trypsin-EDTA solution (0.25%), and were then washed with PBS containing antibiotics and sub-cultured. In the experiments, ovarian follicular cell were cultured in L-15, DMEM/F12 and M199 media, 10%, 15% and 20% of FBS, and were incubated at 20°C, 22°C and 25°C. Cells in L-15 showed better growth but no cell growth observed in the other media. The cells could grow at temperature between 20°C and 22°C, and the results showed that the optimum temperature for cell growth was 22°C. In addition, the growth rate of follicular cells increased when the FBS proportion increased from 10% to 20%, with the optimal growth at the concentrations of 20% FBS. Most proliferation of cells obtained when the L-15 medium was supplemented with 20% FBS and the incubation temperature was 22°C. The most proliferation of cells obtained in L-15 medium with 20% FBS at 22°C.
Nowruzfashkhami M R, Sudagar M, Bahmani M, Salamat N, Mazandrani M. Effect of culture media, fetal bovine serum (FBS) and temperature on ovarian follicular cell in Sterlet sturgeon Acipenser ruthenus Linnaeus, 1758. JAIR 2017; 5 (3) :115-132 URL: http://jair.gonbad.ac.ir/article-1-364-en.html