Study on Peripheral Blood Cell Cultivation and Chromosome Preparation in Stone flounder(Kareius bicoloratus)

Stone flounder(Kareius bicoloratus)was used to explore the condition of the peripheral blood cell cultivation and chromosome preparation.The peripheral blood cell was cultured with RPMI1640 whole culture medium.The conditions of the peripheral blood cell cultivation and chromosome preparation were searched from preservation time of blood,stimulation(PHA and ConA),and so on.The result showed that the cells of whole blood which preserved for 3 days under 4℃ could grow nomally.Under 16℃,both ConA and PHA could facilitate Kareius bicoloratus' peripheral blood cell proliferating,and the best culturing time was 72h.The effect of ConA was better than that of PHA from the shape of hematocyte and the number of leukocyte.Better chromosome samples could be obtained under follow conditions : adding 0.2 ml whole blood and ConA(final concentration 0.1 mg/ml) in 5 ml whole blood medium,cultivating for 72 hours under 16℃,adding colchicines(final concentration 0.6 μg/ml) for 4 hours before cultivation finished,doing hypotonic treatment with 0.075mol/L KCl solution for 35minutes.