We add our drug to the digested cancer cells ,then we add the specific CDs.
before we detect cancer stem cells by flowcytometry.
I think you should determine the proportion of stem cells in the sample before and after using the drug.
Divide the sample in two samples. Then you apply the drug to one of them.
Make the flow cytometry on both samples. The difference between the two samples will tell you if it works on CSC.
it depends on the cells you are using. If you are using breast cancer cells, you can measure CD44hi/CD24lo or ALDH. If colon cancer, you may use CD133. You can comapre their level between treated and untreated.
I agree you should know the proportion of stem cells before treatment for sure. If you have good marker to monitor stem cells in your system the percentage of positive cells for your markers or its level of expression should be a good parameter. If you don't have a good marker you probably should perform a sphere forming assay or a single cell clonogenic in stem cell media, in vitro (testing self-renewal) or test in vivo tumorigenicity at single cells/very low dose (tumor initiation capacity evaluation)...in my opinion of course!good luck!
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