Usually includes the following steps:
1. Choose the appropriate serum
Before replacing fetal bovine serum, it is necessary to select the appropriate serum based on cell type and experimental requirements. For example, some cells have adaptability to specific brands of serum, so when replacing, it is advisable to choose the same brand of serum as much as possible to avoid a decrease in cell growth rate or poor condition.
2. Gradually replace
In order to reduce the stress response of cells caused by environmental changes, it is recommended to adopt a gradual replacement method. The specific operation is to mix the new and old serum in proportion, gradually increasing the proportion of new serum until it is completely replaced with new serum. For example, the new serum can be gradually replaced with the old serum in ratios of 1:3, 1:1, and 3:1.
Suggest the following replacement method:
(1) During the preparation process of the culture medium, mix 25% new serum with 75% original serum and culture for 1-2 passages;
(2) Then mix 50% new serum and 50% original serum for subculture;
(3) Then mix 75% new serum and 25% original serum for subculture;
(4) Finally, culture and passage with new serum.
3. Observe the cellular state
During the replacement process, it is necessary to closely observe the state of the cells. If slow cell growth or poor condition is found, it may be necessary to adjust the replacement plan or re evaluate the suitability of the selected serum.
4. Precautions
When replacing fetal bovine serum, the following points should also be noted:
1.Ensure the quality and consistency of the new and old serum.
2.Avoid repeated freeze-thaw cycles to prevent protein denaturation in serum.
3.Replace cells during logarithmic growth phase to maintain higher cell density.
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