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Effect of culture media, fetal bovine serum (FBS) and temperature on ovarian follicular cell in Sterlet sturgeon Acipenser ruthenus Linnaeus, 1758
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Mohammad Reza Nowruzfashkhami *   , Mohammad Sudagar , Mahmoud Bahmani , Negin Salamat , Mohammad Mazandrani |
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Abstract: (6951 Views) |
| 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. |
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| Keywords: A. ruthenus, Fetal Bovine Serum (FBS), Follicular cells, L-15 |
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Full-Text [PDF 2259 kb]
(2885 Downloads)
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Type of Study: Research |
Subject:
Special
Received: 2016/05/14 | Accepted: 2016/08/20 | Published: 2017/12/21
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