I use DMEM-HG supplemented with 10% FBS, 2 mM L-glutamine, 1% pen/strep, and 2.5 μg/mL fungizone. My cells start to get a weird shape after some passages and the medium starts to have a lot of cell debris. I tried to change the medium and after some hours (like 3-4 fours) it was the same. Then I tried centrifugation, but the cells gained these white spots. However, they are growing well. I thawed another cells (freezed by me when I received the cells from another lab), but after some passages this happens again. I normally wash the cells with PBS before adding trypsin (for 5 min). Do you have any idea of what is happening?
I had a similar problem with the MCF-7 cell line recently. Those spots are vacuoles in the cytoplasm, which indicates that your cells might be exposed to contamination or have a problem with trypsinization timing. If your cells were exposed slightly longer than the usual trypsinization time. Check the trypsinization time for your cell line and use fresh trypsin.
Hello! A few weeks ago I had a similar problem with a breast cell line, you could see a lot of debris and a lot of dots. When performing a PCR for the detection of mycoplasma, we could see a very faint line that suggested that they could be contaminated. To solve it we did 3 things mainly; The first one was to filter the FBS since it comes with a large amount of impurities, then we increased the washings with PBS to 2 and with a duration of 5 minutes in constant movement. Finally, we modified the culture media and increased the amount of Primocin from 100uL to 250uL for almost 2 weeks and we have managed to perceive a decrease in these points. I do not recommend maintaining the primocin increase for more than 2 weeks, as this can affect cell viability. I hope I've helped. Greetings!
It was mycoplasma. Thank you for your answers and help Rodrigo Zañartu Mellado Nilam Parmar
Diana Serenela Salvador ; I am glad to know that my comment was helpful, greetings!
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