In most of the research papers, I noticed that for macrophage polarization of PBMC derived monocytes, the heat-inactivated serum is used during the polarization to M1 or M2. What if I used Non-heat-inactivated serum instead is that fine?
Thank you
Hello Ahmed Al-Omar
The objective of heat inactivation of serum is to destroy complement activity in the serum without affecting the growth-promoting characteristics of the product. Removal of complement activity from the serum is not required for most cell cultures but may be necessary for cultures that are sensitive to the complement activity.
In most cases, heat inactivation does not improve the growth promotion capability of the serum and may actually have adverse effects. This procedure should only be performed if actually required for optimal cell growth.
You may try using non-heat-inactivated serum. May be it will help to improve cell performance. But you need to filter the non-heat-inactivated serum before use to avoid any problems later on during culture.
Hope this helps.
Good Luck!
I agree with Malcolm answer. Heat inactivation is only for inactivating the complement activity.
There are several heat-labile complement protein found in newborn calf serum which may affect the ideal growth conditions of the cells in culture, showing a significant morphological drift. So, heat inactivation is recommended to inactivate the complement system, especially for immunoassays and also to inactivate other undetermined inhibitors of cell growth in culture. With use of FBS, the need for heat inactivation may not be necessary for most cell lines, or minimal unless specified. Moreover, pre-warming FBS to 37°C as most of us do is more than enough to inactivate heat-labile complement components as the levels of these in commercial FBS were just fraction.
However, the most worthy thing to do is to try using both non-heat inactivated and heat inactivated serum for culturing cells of your interest and see the differences.
- Badges
- Science topic
- Similar topics
- Methodology
- Research Methodology
- Research Papers