u/Equal-Race7000

Hi. Looking for some advice re culturing RAW cells. They keep dying on me over the weekends and really hindering my project.
I culture them in non-tc treated 100mm petri dishes as recommended by the lab i received them from. I split every Monday, Wednesday and Friday, seeding 1 or 2 million cells per dish in 10ml.

Media: DMEM high glucose + 10% HI FBS, 1% LGlu + 1% P/S.

Often monday - wednesday and wednesday- friday they are fine, i get up to 15 million cells per dish at 95%+ viability. When i split them on Friday they are fine but by Monday the media has turned yellow and cells are at most 10% viable.

Any advice or suggestions?