细胞培养基本知识（Basic knowledge of cell culture）.docVIP

细胞培养基本知识（Basic knowledge of cell culture） 1 cell culture Cell culture is an important technology in biomedicine. It has been widely used in many fields, such as cell biology, biochemistry, clinical laboratory and so on. The source of 1.1 cells Can come directly from human or animal tissues, or indirectly from the cell line. The former directly from organisms, biological indicators are more close to the in vivo situation; but the extraction process of general cell is complex, and most cells can only be cultured or only 1~2 generation, such as human vascular endothelial cells and mouse nerve cells; cell line benefit is more convenient, can be purchased directly, without separation the process, generally can be stably passaged, but its biological parameters and in vivo have certain differences. Laboratory requirements for 1.2 cell cultures Higher cell cultures are required to allow cells to survive under sterile conditions. A surgical instrument of extracting the cells before each use should be high temperature, high pressure disinfection, with medium water requirements with distilled water and adding double antibody. The used bottles, centrifuge tubes and other utensils are best disposable. Operate on a clean bench in the asepsis room. Before each operation, you should use the UV lamp to illuminate the workbench 20-30min, then wash your hands, change the slippers into the sterile room, and then wash the hands with a new spray. 2 human vascular endothelial cells were used to illustrate cell culture Because of the different nature of cells, the methods of isolation and culture are also different. It is the general source of fresh umbilical umbilical vein (from postpartum within 24h), the specific method is: take the separation of umbilical cord blood (about 25 cm), sterilized in PBS containing penicillin and streptomycin in each 100U/mL, 4C 24h saved startexperiment. 2.1. sampling and primary culture At the ends of umbilical vein respectively inserted into a flattened s