The Establishment Of Cell Culture System And Characteristic Analysis Of Rough-legged Buzzard

Three kinds of tissue cells of Rough-legged Buzzard were obtained by primary explant culture methods in the research. During the process of subculture, biaological characteristics such as cell morphology, growth curve, cell adhesion rate, karyotype were analyzed respectively. All of the Three kinds of tiuuse cells were fibroblast-like growth. Oviduct-derived cells had stronger adherent ability,following by lung-derived cells and trachea-derived cells.The doubling times of lung-derived cells, trachea-derived cells, oviduct-derived cells were29.91±0.39h,33.18±0.21h,30.67±0.28h,with the population doubling times3.54±0.01,4.52±0.02,4.38±0.03respectively/The chromosome numbers of Rough-legged Buzzard was68, within the typical type of ZW.The research have studied the culture condition of cell from lung tissue of Rough-legged Buzzard and the effects of series of factors such as medium, cell detach solution, serum, insulin, inosine on cell culture. During the process of subculture, four kinds of medium (DMEM, DMEM/F12, M199, MEM alpha) were used. The result showed that MEM alpha is the optimal culture medium to the cell growth. Three kinds of cell detach solution (0.25%trypsin,0.02%EDTA,0.25%trypsin-0.02%EDTA) were used for the cell dissociation, and the effect of0.25%trypsin-0.02%EDTA were the best. With serum dependence, the cells fits the medium containing12%serum. Certain concentrations of insulin and inosine promote the cell culture. Four different cryoprotective agents were compared to check out the effect of the cell cryopreservation, and the result showed that10%DMSO+80%DMEM/F12+10%serum is the best.The results above could provide matetials and basis for further research in the field, and the preservation of genetic information of the Rough-legged Buzzard successfully.