Cell Culture From Hepatopancreas And Hemocyte Of Penaeus Japonicus

Mass culture of animal cells is an important area of biotechnology because the cells can be used for virus isolation, vaccine and medicine production. In the present study, firstly, RNA/DNA ratio was used to evaluate three kinds of mediums of different pH. The result showed that pH7. 6 was the optimized condition for hepatopancreatic cells while pH7.2 was the optimized condition for hemocyte of Penacus japonicus. The morphology, structure and ultrastructure of cells were also examined. Secondly, the method about cell culture was improved and the mediums was added extra contents such as the contents of NaC1, MgC12, CaC12, NaHCO and glucase etc. Through the observation of the outer appearance of cultivated cells, it showed that cells grew under the optimal condition of medium added Ca2 and Mg2. If the improved medium was used to cultivate the primary cells from hepatopancreas and hemocyte of Penaeus ]aponicus, single layer of cells were dispersed well and covered 90 percent of area. On this basis, the judgement was on the hepatopancreatic cells in the conditions of different pH, osmotic pressure or addition of Zn2 through RNA/DNA ratio. It showed the optimized conditions to cultivate the cells from hepatopancreas of Penaeus japonicus is pH 7.6, osmotic pressure:630-----730 minol/kg,added Zn2:8Oi,ig/L, compared with the same medium. Thirdly, the effect of metal ions on the cell culture from hepatopancreas of Penaeus japonicus was probed through the RNA/DNA ratio of the cultured cells. The cells grew best when the addition of Ca2was lg/L;the RNA/DNA ratio of cultured cells raised following the increasing of the addition of Mg2the RNA/DNA ratio of cultured cells reached the maximum when the addition of Zn2 was 80iig/L. The cultured cells were promoted sticking to the walls of culture board with the adding of Ca2 and Mg2at the same time. At last, the contents of the general phosphate and inorganic phosphate were determined in the hepatopancreatic cells of Penaeus japonicus cultured and subcultured with the medium added linoleic acid. When the addition of linoleic acid was 16X103M, the concentrations of the general phosphate and organic phosphate reached the maximum. In short, culturing hepatopancreatic cells of Peraeus japonicus needs suitable pH and osmotic pressure, the appropriate measure addition of metal ions, such as Ca2, Mg2 and Zn2, can promote the growth of cultured cells. Mixed addition of Ca2 and Mg2 was beneficial to cells to stick to the walls of culture board. The addition of opportune linoleic acid promoted the growth of cultured cells from hepatopancreas of Penaeus japomz Gus...