I'm trying to do a glucose stimulated insulin secretion assay (GSIS) by adding glucose to MIN6 cells. But strangely the lower dose (3mM) glucose gives a higher insulin secretion than the higher dose (25mM). Any ideas how this can happen?
We are culturing our MIN6 cells with High glucose DMEM (4500mg/dl or 180mM) I've seen people use lower glucose concentrated medium to culture MIN6 cells, could this be the causative factor?
MIN6 are an insulinoma cell line, so I'd expect permanent insullin secretion.
Have you tried to generate a dose-response curve? Could be that at high glucose, the cells simply are overstimulated and reduce secretion.
How well-defined/old (how many total passages) is your clone?
I think that when we use high doses there is a desensitization of the target cells which leads to a weak response
Wolfgang Schechinger and Nadia Chergui, In the original paper the assay is based on, they culture the cells at high glucose. But next time I will try culturing them in low glucose to see what happens.
Also the passage number (was) around 11 when the assay was performed.
I have multiple clones at passage 5-6 in liquid nitrogen at the moment, so I don't think it is because of a high passage number that the cells are not responding properly to glucose stim.
A dose response would indeed be a nice experiment to do.
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