I am trying to do an MTT assay on CSCs (Cancer Stem Cells) but cannot get it work.
Use sensitive fluorescent analogs of MTT. rezasurin is cheap but commercial variant shows better results since contain electron coupling agents and stabilizers (PrestoBlue etc.). also look information about very popular now-days CCK8
As Dr. Daniil have mentioned, Cell Counting Kit-8 (CCK-8) is widely used and have the high sensitivity as compared with MTT assay kit, which depends on the chemical reaction of succinate-tetrazolium reductase.
However, spheroid culture or unattached cells after sorting out for enrichment of cancer stem cells tend to exhibit less response to CCK-8 kit as compared with adherent cells.
What kind of method do you take to get CSCs???
Stem cell tend to have effective system for excretion of xenobiotics. It could reduce efficiency of MTT-test. Try any test with electron coupling agents - it could help to facilitate reduction of dye.
Instead of MTT you can use other method to estimate viability and growth potential in your cell depending of aim (clonogenic assay, spheroid size tracking etc.).
Hi Lixia,
would you please explain why the assay was not working, then it worked?
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