Preliminary Studies On Somatic Cell Iines Construction On For Common Carp(Cyprinus Carpio) And Nuclei-Transfer Techniques For Fish

Common carp and golden carp, both called cyprinidae were chosen for experimental materials in this research. Epithelia of rostral side and fins were taken to culture in vitro. As a result, cells of tail fins were successfully passed from generation to generation, and constructed cell lines. During the culture procedure, the passage cells were observed and their biological characteristics were analyzed, which consist of morphologic analysis, drawing of growth curve, detection of producing rate for clones, insight of adaptation to temperatures, calculation of chromosomes and investigation for cells frozen to thaw. Meanwhile, subcultural cells regarded as donors were transferred into unfertilized eggs with nuclear of common carp and golden carp, the results are as follows:Cell lines from rostral side and caudal fin of common carp were respectively established successfully, and passed to 65 generations at present.Biological characteristics were analyzed for primary and secondary cultural cellsMorphologic observation. There is an inclinable increase for spindle-shaped cells during cell culture.As the growth curve described, subcultural cells grow best between 48h and 72h, and divide very faster.The data calculated for clone forming rates demonstrate that clone forming rates of the 31st cells are 70% more.Temperature adaptation experiments delicate that the cells from common carp have a strong ability to tolerate temperature, especially the ability to tolerate lower temperature is greatly stronger than to higher temperature.Number of cell chromosomes pronounce that both primary and secondary cell chromosomes are 2n=100; belong to normal diploid, and haven't any variances.Two methods are used to cryopreserve – revive cells. One is generally deducing temperature by using different refrigerators and preserves cells with DMSO; alternatively, by depending the sample in the hole of liquid nitrogen vat and preserves cells with glycerol. The cells are calculated before cryopreserving and after reviving, the data show that the reviving rate of cells preserved by the former method is slightly higher than that of the latter one, and there isn't any change in cell morphology. 3. The subcultural 4-10 generational cells as donors were transferred into unfertilized eggs with nuclei from common carp and golden carp as receptors, finally, the embryo developed to gastrula stage.