The Cell Culture Of Different Tissues From Chicken Embryo And The Immortalization

The chicken embryo cell line with immortalization can be used in the diagnosis and testing in the etiology and pathology of avian, the production of bacterin and the test of its titer and safety. Considering no such cell line recently, the research cultured numbers of tissues from chicken embryo in vitro, and make some study of their basic biological characteristics. Some methods as physical,chemistrical and cell fusion are used to induce the different cells from chicken embryo, aiming at a chicken embryo cell line with stable physiological and growth.The research cultures three kinds of cells in vitro, the body tissue cells(BTC),the heart tissue cells(HTC) and lung tissue cells(LTC).And keep the cells in liquid trogen as cell resource.Then,we did some complete analysis of biological characteristics of those cells above. The synthetical analysis of cells includes their morphologic shape, the growth curve, the division index and the analysisof karyotype.With the method of curettement and separation for phase different, we got corresponding cells from three tissues hereinbefore, with,same size, shape and growth characteristic. A further identify of form and the check of pollution with mycoplasma were conducted with the dyeing methods of DAPi. Using the monoclonal antibodies against Vimentin, Desmin,Cytokeratin-19,the identify of immunocytochemistry was proceeded wih the body tissue cells(BTC),the heart tissue cells(HTC) and lung tissue cells(LTC),and BTC is fibroblast cell, HTC is myoblast cell and LTC is epithelial cell.The research usd MNNG, sodium azide and UV to induce the third,fourth passage cells from different tissues, LD50 calculated,then select the cells after inducement, subculture over a long period of time. And two lines(BTC,HTC) subculture to the 36th and 29th.The research seeks the best condition of cell fusion between the cancer cells (SP2/0 and hePG2-) and the cell from chicken embryo,using PEG as fusogenic agent. The fusion cells were selected in the selective medium of HAT, and we got cells of hybridoma.After cell clone and subculture,the speed of hybridoma cells accelerates apparently.