Study On The Technology Of Transgenic Chicken Using Blastodermal Cells Medias

Adopting cationic liposome transfection and microinjection technique, we preliminary established transgenic chicken model. The donors we used were blastodermal cells which carry enhanced green fluorescent protein (EGFP) gene. The research is to establish a technical platform for using transgenic chicken as bioreactor. The main research results are as follows:1. When culturing blastodermal cells of chicken in vitro, the cell attachment rate is maximum with 10% fetal calf serum. The differences are insignificant by using two different media M199 and EMDM. The cell attachment rate will reach the highest after culturing 24 hours in vitro. The highest cell attachment rate is about 60%.2. When transfecting the report Gene to blastodermal cells of chicken in vitro, there are about 60% transient expression both for adherent cells and suspension cells. The transducting efficiency of White Leghorns primary blastodermal cells is highest when the liposome amount is 4μg and plasmid DNA amount 3μg.3. During microinjecting on the recipient eggs, the hatching rate of eggs which are windowed on equatorial plane is the highest, reaching 26. 74%. The hatchability with egg-shell membrane and sealing-glue combination sealing is the highest, reaching 27.08%. The hatchability has significant differences between no windowing-processed and windowing-processed recipient eggs (p<0.05). Because the structure of the recipient eggs which had been windowing-processed was damaged, their hatchability reduced significantly. And the hatchability of recipient eggs microinjected blastodermal cells is 28.43%.4. Detecting the transgenic chicken embryos and the first weekly born chicken by fluorescence and PCR, we found there is weak green...