How to Digest Cells from Cell Culture Flasks

 Cell culture flasks are mostly used for the cultivation of adherent cells. Different from the growth of suspension cells in the culture medium, adherent cells need to grow on the surface of the container, so the cells must be digested before harvesting or passage. There are many ways to digest cells, the common ones are as follows:

1. Enzyme digestion

This is also a method that we use more in our daily life. Generally, trypsin is used for digestion, and the concentration is between 0.25% and 0.5%. The digestion time varies according to the cell type, operation method, temperature and other factors. After digestion, it is necessary to stop the digestion with serum or serum-containing medium.

T25 Flask

2. Physical method

Without the aid of trypsin, directly pipette or use a cell scraper to digest the cells on the surface of the cell culture flask. This method can be used for cells that are firmly attached. However, it should be noted that pipetting and scraping will cause certain damage to the cells, and it is generally not recommended to use this method.

3. Freezing method

Using the principle of cell shrinkage after freezing, the cells are detached from the culture flask. This method causes little damage to the cells, and is suitable for cells that are not particularly tightly attached but are particularly delicate. Such as mesenchymal stem cells, DC cells, etc.

T225 Flask

4. Ion chelating agent

Does not destroy cell surface molecules, only chelates with CAMs. Generally, more EDTA is used, and the concentration is about 0.02%. It acts on cells and interstitium, and also has a certain effect on intercellular space. Note that it can significantly affect pH and is only soluble in slightly alkaline conditions. Therefore, the pH should be adjusted during preparation. It cannot be finalized. Therefore, the digested cells should be washed again.

The above are several methods commonly used to digest cells in cell culture flasks. On the whole, enzymatic digestion is the most widely used method. Of course, the appropriate digestion method should be selected according to the type and characteristics of the cells.