Hello everyone I need help,
I am working with KGN granulosa cell line (DMEM/F12 media, 10% FBS, 1% Pen/Strep)
I am treating the cells with PMA at 100nM concentration to activate PKC, but everytime I add PMA my cells are shrinking and look like they are dying.. Does anyone have ever experienced something similar ? Any idea why they are looking like that and why does the PMA would induce that ? (I also tried with lower concentrations; 20. and 40nm and had the same results..).
Thank you so much
Hello Fortin,
If you have the chance I would suggest checking the water levels and the CO2 infusion of your incubator, I have noticed that even though sometimes not apparent slide changes on these basic conditions can hit hard your culturing.
Alternatively, you can slowly increase the % of FBS and see if they can come back to how they are supposed to look like.
Hope it helps,
Marcos
Thank you very much, I’ve checked and CO2 have been constant at 5%, water is ok too! I’ve noticed that after 16-24h they start looking normal again.. but I’m still worried because ultimately I will treat them for only 12h and so the results will be skewed by this problem..
Chloe
PMA can induce cell cycle arrest, apoptosis or non-apoptotic cell death in some cell lines. The effect depends on cell context. For instance, in LNCaP cells, treatment with PMA induces PKC-dependent apoptosis through activation of JNK, but in AsPC1 cells this treatment induces non-apoptotic and PKC-independent cell death.
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