Hello,
I'm having a trouble with HaCat culturing from stock (the medium for stock with the ratio 9 full DMEM : 3 FBS : 1 DMSO). The cells seem to secret many black dots as shown in the picture and their morphology are abnormal, too. I tried to subculture over several passage and wash carefully but the black dots still appear. Does anyone have the same trouble like me? Please give me some advices. Thank you!
P/s: I also wonder if the black dots are mycoplasma, however they do not move and seed to the plates.
Hi Nguyen!
Did you solve the problem? I also encountered those small adherent black dots on my HaCaT culture dish and the cell condition seemed to be affected. Just don't know how to eliminate them. Thank you.
Hi Huidong Liu The phenomenon is caused by cell senescence. I tried to use HaCaT with the lower passages. Currently, my HaCaT cells are being cultured in DMEM 10% FBS and 1% AB (all were bought from Gibco). Last time my cells were cultured in DMEM (Hyclone Co.) and FBS (Hyclone Co.). So, I was observed that cells condition becomes better with less black dots.
Did this cell condition affect your experiment result? I am studying inflammation on HaCaT cell with LPS but LPS cannot induce inflammation.
Hi everybody,
I also had the same observation with haCAT cells and trying to figureout the reason whya nd how to fix it.
Amal.
Hi everyone! I am facing a similar issue with the HaCat cells. Upon revival from the stock, the cells seem to secrete these black dots, and they tend to gradually increase in number with every passage, despite carefull washing everytime! I don't have this issue with the cells under culture currently. This only happens everytime I revive a stock. It it anything to do with the cell line as such? Because, this phenomenon does not occur when we try with other cell lines like RAW, or FLS.
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