I am culturing transfected Drosophila S2 cells. I would like to culture them into a serum-free media (SFM). However, I have read from a paper that cells cultured in SFM are more sensitive to antibiotics (including the selection agent).
So with this, is it recommended that I stop adding antibiotics or I decrease the antibiotic concentration several folds?
Thanks
I have no direct experience with this yet, but I looked (hard) for this information and found that some people DO add selection antibiotics to serum-free medium. See for example this paper: Zitzmann et al, Biotechnology Reports 19 (2018) e00272.
I am leaning toward the feeder layer single cell cloning procedure described in this paper (also see Yamashita et al Protein Science 2017 26:2291 for a different cloning method, soft agar) as two of my lines seem to have lost expression of my protein during the six weeks after transfection.
I also found this paper in which antibiotics are added to SFM (but in the Notes they say it is not necessary-?!)
https://link.springer.com/content/pdf/10.1007%2F978-1-4939-3043-2_17.pdf
I can now report that my S2 cells are growing fine in shake flasks in serum-free medium with 2 ug/ml puromycin.
Thanks for the resources! I used blasticidin as a selection agent for my cells shaken at around 120 rpm with 0.1% Pluronic F-68 in serum-free medium. I followed the recommendation of Corning and the cells grew well and seemed to have expressed my protein.
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