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The chick embryo immortalized cell lines can be used to pathological diagnosis, vaccine production , and its efficacy and safety testing . In view of the domestic establishment of the cell lines , this study by in vitro culture of chick embryo tissue cells , the basic biological characteristics were determined , and the use of physical, chemical , and cell fusion method , in vitro induction of chick embryo tissue cells conversion to occur , the create the chick embryo immortalized cell lines . Take the chick embryo body -derived cells ( Body Tissue Cells BTC ) study , cardiac -derived cells ( Heart Tissue Cells , HTC ) , lung -derived cells ( Lung Tissue Cells , LTC ) in vitro and cryopreservation . Indicators conducted a comprehensive analysis of the morphological characteristics of the cells , the growth curve , mitotic index , chromosome karyotype . Using the the curettage method and the difference of digestion , separation and purification of the corresponding tissue cells from the cells of the three organizations . The purified cell size is similar to the morphologically similar growth characteristics . DAPI staining of cells isolated and purified further morphological identification of mycoplasma contamination detection . With anti-vimentin monoclonal antibody , anti- desmin monoclonal antibody , anti-keratin -19 monoclonal antibody BTC , HTC , LTC cells do Immunohistochemical identification , detection came to the BTC fibroblasts , HTC into muscle cells , LTC epithelial cell types . MNNG , sodium azide and UV-induced third and fourth generation of the different organizations of the chick embryo cells , to calculate the LD50 . Screening of the induced cells , and long-term subculture and cryopreservation, of which two ( BTC , HTC ) cells were passaged to 36 and 29 generations . Explore different molecular weights , different concentrations of PEG for fusion bonding agent in the HAT selection medium under the optimal conditions for induction of cancer cells and chick somatic cell fusion , screening the fused cells to obtain hybridoma cells . Cloned and subcultured after hybridization cell growth rate was significantly improved.
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