They are cultured with DMEM supplemented with 10% FBS. What could be the reason for the layer formation? However, a persistent layer was observed forming on top of the culture the very next day, despite daily PBS washes. Even after supplementing the medium with additional amount of gentamycin, no significant improvement was noted. What could be the possible cause of this layer formation, and what steps can be taken to eliminate it?
Hi Senthamizh Gopal,
Although you haven't mentioned the kind of thing forming the layer, it seems to be mycoplasma contamination in the culture. U87 cells are very susceptible to mycoplasma contamination. They proliferate overnight and aren't really bothered by PBS wash or the addition of fresh media. Neither gentamycin can help in this case. The only way is to throw the culture, find the source of contamination, and start afresh.
Best wishes.
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