Dear all,
I have been having trouble with EdU staining of primary human meningioma cells. I am finding that even with 24 hours of incubation with EdU reagent, I am seeing a very low (0-5%) EdU staining percentage in untreated cells. Unusually, this % dramatically increases with increasing concentration of the drug I am testing. I have seen this across four separate tumours now.
I attempted to clarify if the drug was really increasing proliferation, which would be the opposite of the intended effect, by double-staining with both EdU and Ki-67. The Ki-67 looked as I would have hoped, with a starting positivity % of about 40%, decreasing to 15-20% with the use of the drug. EdU again, started at almost nothing in the untreated group and raised to around 20% at the highest drug concentration.
Has anybody seen this pattern before or might be able to explain it? I am aware that Ki-67 and EdU indicate different parts of the cell cycle, and that Ki-67 is usually higher, but I wouldn't expect to see completely opposite patterns. EdU has been reliable for all of my other work with this drug in immortalized cell lines thus far with only 4 hours exposure to the reagent; it is only primary human cells that have posed this issue.
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