I seeded 2x10^6 RAW264.7 cells in T-75 flask. Normally, they will have a confluency of 90-100% within 2 days (no change in cell culture media color). However, recently, the cell culture media in the flask turns yellow within 2 days, with around 60-70% confluency, along with an abnormally large number of floating cells. The shape of the adhered cells seems to be normal. Usually, RAW264.7 cells do not float unless they exceed 100% confluency, but my cells have not yet reached 100% confluency and they already float. The viability of the cells is 70-85%. I checked for mycoplasma, and results were positive. From researching various sources, mycoplasma contamination usually does not affect media color this much.

Does anyone have previous experience with mycoplasma contamination in RAW264.7 cells? Could mycoplasma be the only factor in all of this, or is there something else.

Thank you in advance.