I want to culture microglia/astrocytes and stimulate them with LPS and Poly I:C. Then I am planning on taking this stimulated media and transferring it to a different cell line. Is there any way to flash freeze the microglia/astrocyte stimulated media so I can use for later experiments? Has anyone tried this?
I used to work with conditioned media from fibroblast to treat epithelial cells. We centrifuged (to eliminate contaminating cells) and freeze conditioned media for future uses. We didn´t have problems with that. Of course, it is not the same as fresh media but most of the components of the conditioned media such as grow factors or proteins can be freeze. Freeze-thaw cycles are not advised. If you are planning to freeze it, separate it into aliquots before freezing.
Hi Christina,
I've done a similar experiment with conditioned media before. I stimulated primary murine cortical microglia, microglia astrocyte co-cultures and astrocyte cell cultures with LPS, INFy and ATP and froze the conditioned media. We found that already after one day the induced inflammatory response of the conditioned medium was weaker compared to the fresh conditioned medium.
Therefore, we tend to use fresh conditioned media. Of course, freezing media and using it later is just very practical. Depending on your readout it may be that it doesn't matter for your attempt to freeze your medium.
cheers,
Pardes
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