What are the benefits and disadvantages of using heat-inactivated FBS in cell culture?
- Heating inactivates complement. Active complement can participate in cytolytic events, contract smooth muscle, release histamine from mast cells and platelets, and activate lymphocytic and macrophage cells. Applications where heat-inactivated serum is recommended include immunological studies and culturing of embryonic stem cells (ESCs), insect cells, and smooth muscle cells.
- Heat inactivation helps to achieve bottle-to-bottle and lot-to-lot stability by neutralizing many factors that can vary largely from lot to lot.
- There aren't necessarily disadvantages to heat inactivation of FBS, but there is some evidence that suggests there may be no added benefit to it unless you are carrying out immune studies.
Note: Heat inactivation is performed in a 56 degrees C water bath for 30 min with swirling every 10 min or so for heat distribution and to lower the degree of protein aggregation/flocculant precipitation. Note: If the time or temperature is exceeded, the serum may thicken to a gel. If this occurs, the serum is no longer usable. Unnecessary heat inactivation can take up time and potentially lead to wasted reagents if a mistake is made during the protocol1.
1. Pellerin, et al., Bioengineering, published in 2021.
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When should I use heat-inactivated serum?
Heat-inactivation of serum inactivates the “complements” in the serum. In general, it is not necessary to use heat-inactivated serum to culture most cell types. We recommend using heat-inactivated serum when working with immune type cells or for immunological applications.
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I would like to order heat-inactivated FBS. What conditions do you use for heat-inactivation of FBS during the manufacturing process?
Serum is heat inactivated by heating at 56 degrees C in a water bath for 30 min, swirling the bottle approximately every 10 minutes.
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What is the shelf life of Fetal Bovine Serum, qualified, United States once thawed? Can I refreeze it after thawing?
The stability after opening and thawing is application-dependent and most often, it is used within a week or less. The product can be thawed and refrozen. This is standard practice for making aliquots. We recommend that you make smaller aliquots and use as needed.
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What is the difference between "qualified" versus "certified" purity grade FBS?
The main difference is in the Quality Control (QC) test specifications. “Qualified” and “Certified” FBS go through the same QC tests but “Certified” FBS has additional QC tests as well as more stringent QC test specifications for endotoxin and hemoglobin levels. The additional QC tests in “Certified” FBS include biochemical and hormonal profiling.
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