I am not sure why one shouldn't add serum to the media that we add to cell cultures to obtain conditioned media the following day. Could someone help me get up to speed with this? Thanks.
When you add serum, you are introducing some unknown biological factors (i.e cytokines, proteins, etc). So, you cannot say that the effect that your are observing is only related to the soluble factors that supposed to be there. But, if you use the media+serum (the one that you used to obtain the cm) as negative control, you can discard the serum´s effect on your experimental paradigm.
Serum is a unspecified mixture that will effect your experiment without any clear control over the events. By adding serum your conditioned media will become tainted with factors that are not part of the conditioned media.
Hi. I think It depends on your chemical/ protein/cytokine that you are going to add in the media. Serum have multiple proteins/factors, and might some of them have binding capacity with your agent. This can effect your results. Otherwise as Litia said, serum plus media can be used as a negative control.
Further to the other valid points, depending on what you're conditioning the medium with – if, for example, a biomaterial is the conditioning agent, or a carrier is releasing a drug – the proteins in the serum may alter the material's / carrier's release or degradation kinetics.
Thanks all. I was also wondering whether one can freeze a cell pellet with small amounts of DMEM on top of the pellet. This is the residual DMEM that one cannot aspirate out of the fear of dislodging the pellet. Also, the storage is at - 80 C for about 40 hours and the cells will be processed for miRNA extraction. The cell line I am using is the murine C2C12.
- Badges
- Science method