In vitro investigation on response of cultivated hepatocytes from Klunzinger s mullet, Liza klunzingeri, to temperature and fetal bovine serum

The appropriate temperature and growth supplementary factors such as fetal bovine serum (FBS) are the necessities needed for cell growth in culture medium. In this regards, the effect of Leibovitz15 (L15) medium containing different concentrations of fetal bovine serum (FBS) was assessed on growth and proliferation of cultivated liver cells of klunzingerchr ( 39 )s mullet, Liza klunzingeri, incubated at various temperatures. Ten fish were dissected and the liver was removed and cut into small pieces. The hepatocytes were then disconnected using collagenase type IV. The cells were cultivated for 2 weeks in L15 containing 0, 5, 10 and 20% FBS and incubated at 20, 25, 28 and 30 degrees centigrade. According to the results, an increase in temperature up to 30 °C led to an increase in growth and number of cultured cells, but temperature rise above 30 °C had no significant effect on growth of cultivated liver cells from L. klunzingeri. Cells cultivated at 20 °C did not attach to the culture plate and died. Growth rate and the number of cultivated hepatocytes increased dose dependently and the maximum amount of growth was recorded in cells treated with 20% FBS at the end of the experiment. In conclusion, the general conditions for primary culture of hepatocytes from L. klunzingeri reported in the present study represented a foundation to establish an in vitro system for L. klunzingeri liver cell culture.