I've cultured some C3H10T1/2 cells in 24-well plates for 72 hours. Cells are maintained with Basal Medium Eagle supplemented with 10% FBS, 1% L-Glutamin and 1% Penicillin/Streptomycin. Medium was changed after 2 days of culture, and cells were wash with 1X PBS prior to adding fresh medium. On the 3rd day of culture, I observed that cells in some wells looked like dead cells as they weren't bright, their cytoplasm seemed very clear, and they had weird morphology compared with healthy cells in other wells or plates.
I'm not sure if they're undergoing cell death or apoptosis. I couldn't think of any possibilities since cells in other wells of the same plate look normal, and all cells are treated in the same conditions.
(The first 2 photos are abnormal cells. The 3rd one is normal)
TUNEL is not specific...how about using electron microscopy along with MTT assay? also what about the annexin-PS test?
Simply add Tripan Blue and see if they are blue or not. If they ar blue, they are dead, an then you can analyze the way they died (apoptosis, autophagy, necrosis, etc). Other way you can do is, if you have the possibility of using fluorescence microscopy, to stain the cells with Propidium
Hi Mizan, Ahmed, Jacquesm Alberto, thank you so much for your suggestions!
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