I have problem about MTT assay in U87 cell line. After add MTT solution in U87 cell line, the cell were floated. Who used to find the same problem. Why is cause of this action? Please answer and suggest me for solving of the problem. Thank you.
I'm not certain about this, but years ago I asked all my students/researchers to be careful of the 'type' of plastic they were incubating cells on. We had problems when researchers used 'cheaper' plastics. Now everyone uses the more expensive plastics with better 'cell adhesion' characteristics. One other factor I noted what the students were quite harsh when the were washing/draining and handling the cells. They increased the cell adhesion by handling the cells very gently when they were changing media and reducing the time the MTT was incontact.
Can you do a titration of the MTT? i.e. Try 1:2 dilution or a 1:4 dilution to see if it helps. My instincts are however it could be the plastic and the U87 cell haven't quite 'stuck-down'?
Hello Chonthicha Jantanon
I too faced a similar problem but with the cells in the side wells of a 96-well plate, which detached with complete layer after some time while the cells in the middle wells stayed attached.
Later I found a solution to this problem. It was because of the 'Edge Effect'. Temperature difference across the plate and evaporation effects in the side wells during incubation was the cause of such type of cell behavior. In the side wells, because of evaporation, salts from the culture medium accumulate in the wells affecting cell attachment and metabolism.
If you are experiencing cell detachment for the entire 96-well plate, then I would suggest you work gently with these cells at all times as vigorous or harsh handling of cells can result in cell detachment and damage or even cell death. Never pipette medium or wash buffer directly onto cells, always add it gently to the side of the well to avoid harming the cells.
Best.
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