علوم زیستی دریا
Negin Derakhshesh; Abdolali Movahedinia; Negin Salamat; Mahmoud Hashemitabar; Vahid Bayati
Abstract
The liver is one of the most important organs of animals that know as a detoxifying and filtering toxin in the body and the most studies was used the target organ. In peresent study, hepatocytes were isolated from 8 immature grouper fish (equal to the number of male and female). Then, anesthetized fish ...
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The liver is one of the most important organs of animals that know as a detoxifying and filtering toxin in the body and the most studies was used the target organ. In peresent study, hepatocytes were isolated from 8 immature grouper fish (equal to the number of male and female). Then, anesthetized fish and the head was separated from the body. The fish wiped with 70% etanol alchol. The liver was removed was cuted with scissors. Finally, the cells digested by collagen type 4. Then, the cells incubated in 30^C and 20% FBS+ITS during two weeks. The doses of pollutant (100 µM, 2*100 µM, 3*100 µM) added to samples, then were studied in 0, 6, 12 and 24 time. The morphological analysis exhibited that the cell death incubated by B[a]P polluted by concentration of 100 µm results in 50% of apoptosis cell death. Our finding from this study showed that using lower dosage of B[a]P during 24h treatment period cause inhibition of proliferation in time and dose dependent manner and higher doasage cause lysis and necrosis.
علوم زیستی دریا
negin derakhshesh; Abdolali Movahedinia; n s; m h; v b
Abstract
Liver is one of the most important organs in vertebrates that have important roles in detoxifying. The main purpose of the present study was developing appropriate methodology for the primary cultivation of hepatic cells from orange-spotted Grouper, Epinephelus coioides. In present study, hepatocytes ...
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Liver is one of the most important organs in vertebrates that have important roles in detoxifying. The main purpose of the present study was developing appropriate methodology for the primary cultivation of hepatic cells from orange-spotted Grouper, Epinephelus coioides. In present study, hepatocytes were isolated from five grouper individuals. Initially, the fish wiped with 70% ethanol. Liver were removed and cut into small pieces with scissors and hepatocytes were disconnected using different enzymatic digestion with collagenase (Type 1 and 4) and trypsin and additional nutrient materials in culturing mediums. Then, cells were cultured for 2 weeks in Lebowitz L-15 under 3 methods: 1. Using enzymatic digestion by trypsin, 2. Using enzymatic digestion by collagenase (type 1 and 4) and 3. Using nutrients and additives was cultured. Finally, effects of different incubation temperature (20, 25, 28, 30 and 32 degree of Celsius) and Bovine serum content (0, 10 and 20% and 20%+ITS) on cell growth were estimated. According to the results, digestion by collagenase type 4, resulted in more cell colonization and growth in comparison with other methods.In conclusion, the best culture method for primary hepatocyte from orange-spotted Grouper, Epinephelus coioides., was using ITS+20%FBS under 30 degree of Celsius incubation temprature.