细胞培养作业（Cell culture operation）.doc

细胞培养作业（Cell culture operation） I. including tissue culture, cell culture and organ culture. Tissue culture technology: refers to the organization simulated physiological environment, removed from a body in sterile, appropriate temperature and certain nutritional condition, of the structure and function of the survival and growth and to maintain its methods. (1) pollution-free environment Non-toxic and aseptic culture is the primary condition for ensuring the survival of cells. (2) suitable temperature Cells in the body of the optimum temperature is 36.5 ℃ + 0.5 ℃, deviation, can affect cell metabolism, and even death (3) gas and pH value O produces energy to supply cells to grow, multiply, and synthesize various components. Most cells do not have enough oxygen to survive. CO2 is both a product of cell metabolism and a component of the cell, which is used to maintain the pH value of the medium, and the cell requires 7.2-7.4. (4) nutrients: sugar, inorganic salts, amino acids, vitamins, and cell growth factors (5) osmotic pressure (infiltration unit) Ion strength: 260-320 m Osm/L medium Primary cell: the first culture of cells, tissues and organs removed directly from the body. The cell line of cell population that has proliferative capacity from the first generation of cultured cells. Diploid cell: the number of chromosomes in the cell group has the same or the same number of cells as the original donor. Cloning cell lines: cell lines formed by individual cell proliferation from a biological identified cell line using single cell culture or screening methods. To get a large number of cells, the existing tissue should be fully dispersed, allowing the cells to dissociation to form a single cell. If blood, amniotic fluid, chest water or ascites are used centrifugation method to obtain. The specific methods are as follows: (1) separation of suspended cells Material: blood, ascites, chest water Centrifugal method: the simplest method is to use a low speed of 1000r/min for