动物细胞培养和核移植技术课程.pptVIP

Animal cell culture and nuclear transfer technology Introduction about animal cell culture 1.The process of culturing animal cells The individual cells quickly attach to the sidewall in the suspension, which is known as the adherent cells. When culturing attached cells, cells must attach the substrate to the growth and proliferation, which requires culture bottles or Petri dish within a smooth surface, non-toxic, and easy to attach. Then if cells begin to mitosis, the number of cells grows fast, and when the adherent cells grow to the surface and contact with each other, the cells would stop proliferation. The phenomenon is known as contact inhibition of cells. People usually name the initial digestion of animal tissue culture as the primary culture. Cells covering over the sidewall need the trypsin to retreat, and then divided them into bottles to continue to foster and proliferate. This training process is usually called a subculture. Subcultured cells are not easy to pass on after spreading 10 generations. In general, the cells that spread to about 10 to 50 generations, the proliferation gradually slows and completely stops, at that time, the karyotype of some cells could mutate. When continually subcultured, a small number of cells can overcome the natural limit of the cell life, and access to the immortal nature, so these cells have mutated and are moving in the direction that are equal to the cancer cells. Currently in use or cryopreserved normal cells are usually less than 10 generations, in order to maintain the cells in a normal diploid karyotype. The conditions of culturing animal cells In general, animal cells cultured in vitro require the following conditions. Sterile and non-toxic environment is important.： We should firstly ensure that the cells were cultured in a sterile and non-toxic environment, which means applying the sterile medium and cultured appliances. Usually we also need to add a certain amount of antibiotics in cell culture medi