I grow LC-540 Leydig cells, the problem with me is as shown in the picture a. The ones rounded in yellow circles are black particles attached to the plate loosely it does not show any movement, but once i wash those cells with PBS it gets removed off easily as shown in picture b. After subculture i get back these particles.
But i feel like these are dead cell particles and i am not sure if this is contamination too..
Please help me identify the problem and need suggestion to get rid of these.
Hello,
Firstly do you use any magnetic beads to isolate the cells by using (MACS)?- magnetic assisted cell sorting. If not it may be mycoplasma infection. There are ways to detect that whether its true or not.
This doesn't look good and might be contamination. I suggest you check the whole incubator and ask if anybody else using this incubator has the same problem with their cells, if not - discard this dish and start new culture.
It looks like a Mycoplasma contamination. As Yulia says, I also recommend you to start with new cells.
Thanks yulina and Maria if I did start with new cells and it turns out that after 3 or 4 passages it's back to this stage.
better u can use 0.8 % trypsin EDTA and find the suitable culture plate from NEST company. Thts it.
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