The process and precautions for harvesting cells in the cell factory

With the rapid development of industries such as biopharmaceuticals, vaccines, and monoclonal antibodies, the importance of cell factories in large-scale cell culture has become increasingly prominent. Cell culture is a rigorous work, with strict control over the cell growth environment, the addition of culture media, and the harvest of cells.

When using the cell factory to harvest cells, the following procedures must be strictly followed:

1. Pour out the culture solution after incubation, and wash it with calcium-free and magnesium-free phosphate buffer (CMF-PBS) (40-50 ml/layer), and repeat the washing if necessary.

2. Digestion: The digestion solution (10-40 ml/layer) is preheated in advance.

3. Collection: Centrifuge at 1000 rpm for 5 minutes to remove the digestion solution and collect the cells.

4. Washing: Wash the digested incubator with CMF-PBS or culture medium.

Pay attention to the following points when harvesting cells:

1. Make sure that CMF-PBS is completely immersed in each culture surface, and gently shake the cell factory back and forth to clean the remaining culture fluid.

2. Digestion solution is evenly distributed to each layer. Gently tilt the incubator back and forth to ensure that the digestion solution is completely immersed in the culture surface. Gently tap the incubator to help the cells detach from the surface.

3. For the cell factory, since the digestion state of the cells in the middle layer cannot be clearly observed, it is recommended to refer to the digestion state of the culture flask or monolayer cell factory under the same culture conditions. You can also use a dedicated multi-layer cell culture observation platform to observe the growth status of each layer of cells.

3. If there are a large number of cells in the cleaning solution or the culture layer of the cell factory, it is necessary to clean multiple times or adjust the operation of cell digestion.

4. A slight deviation of the culture temperature may affect the harvest rate of the cells, so pay close attention to the culture temperature is the set temperature.

The above is the process and precautions for harvesting cells in the cell factory. Cells are very sensitive to the environment. Doing a good job in cell harvesting is the key to the success of cell culture, and all links need to be strictly controlled.