PBMC's - (Mar/19/2008 )

Hello everybody,

Can anybody help me figure out what went wrong?

PBMCs were in GM-SCF and IL-4 for 5 days, on 6th day when i wanted to stimulate them, wells looked yellow.
there's was no contamination. i didnt want to continue so i seperated two more donors, and these two plates in incubation for 5 days. on 6th day one of the plate appeared yellow and no contamination. plz help me what could be the problem. medium is pink in color normally the wells look pink till i collect the supernatants for cytokine analysis.


Carry

Hi, sorry if it's a stupid question but have you checked your cell density? Do your cells look normal or activated?
Do you change your media at all before day 6? It sounds to me like your cells might be growing to much, consuming nutrients, and changing the media pH.

I agree with almost a doctor,
what do you mean bu yellow?
if the medium color change to yellow, that mean the cell is growing (Ph change the color of fenol red)

Ya i think maybe your cells are too confluent. From my experience, the media would turn yellowish after culturing PBMC (with a starting cell concentration of 1x10⁶ cells/ml in 24 well plate) for 4 -5 days. I think you should check and see if there are too many cells in your culture.