1. Microcarrier selection: Firstly, using three small microcarriers for culture experiments, it is better to observe the cell sorption rate and the number of cells in a certain period of time to obtain the largest amount of cells.
2. Hydration: Weigh a certain amount of microcarriers into a container, add 50-100ml per gram of microcarriers, add Ca2+ and Mg2+-free phosphate buffer (PBS), and place at room temperature for not less than 3 hours. From time to time, gently agitate and then wash once with fresh PBS.
3. Disinfection: It can be sterilized by high pressure steam, or immersed in 70% alcohol after hydration, and then rinsed once with sterile PBS.
4. Subculture: When the microcarrier culture is continuously carried out, the cells can be separated from the microcarriers, and the microcarriers carrying the cells and the new microcarriers can be mixed and cultured to move the cells to the new carrier. If other experiments are performed or when it is necessary to isolate the cells for subculture, as in the conventional culture, the cells are first detached from the surface of the microcarriers by the action of EDTA + trypsin solution. After the cells are detached from the microcarriers, the cells can be self-sinked at the bottom by natural sedimentation, that is, at room temperature for 5 minutes. The cells are mostly in the supernatant, and then the supernatant is centrifuged to obtain cells. If a higher degree of separation is required, it is filtered with a nylon mesh or stainless steel mesh having a pore size of 100 μm, and then centrifuged to obtain a relatively pure cell.
Soup Plate ,Soup Plate Bowl,Bowl Soup Plate,White Soup Plates
Guangzhou Concept Home Ltd , https://www.concepthomelimited.com