If you want to show inhibitory effect of some drugs on cancer stem cells , is it sufficient to isolate cancer stem cell from clinical specimen depend on specific cell marker that previously test by re-population assay , or its mandatory to do re-population assay again ?
I read that gold standard measure of a stem cell is maintenance of long-term clonal growth in functional re-population assays, however to what extent are other methods used to isolate cancer stem cells are efficient ?
Hello
See this links & attachment will help you
Good Luck
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0079798
http://www.nature.com/stemcells/2009/0903/090326/full/stemcells.2009.47.html
Clonogenic assay is the best functional assay to analyze cancer stem cell growth and targeting. Indeed its mandatory to do a secondary spheroid analysis to make sure that all cells are the clonal expansion of cancer stem cells. Apart from that xenograft is best to utilize the function of cancer stem cells.
I would recommend both clonogenic assay and in vivo tumorigenesis assay as Chandrakesan mentioned above. Since you are testing drugs, you may also test sphere formation after apoptosis. For example, you may treat the cells for 24 hours and collect the floating cells and replate in new medium and look for colonies after 4-16 hours or at both time points. If you see spheroids at these times, that indicates the presence of cancer stem cells (meaning your drug cannot kill cancer stem cells completely). As a positive control you may use cisplatin_lithium as described in my paper (link below). If you don't see sphereoids in positive control, then the interpretation can be the cancer stem cells in your cell type may not be able to survive after apoptosis.
https://www.researchgate.net/publication/233742776_Blebbishields_the_Emergency_Program_for_Cancer_Stem_Cells_Sphere_Formation_and_Tumorigenesis_after_Apoptosis
Article Blebbishields, the Emergency Program for Cancer Stem Cells: ...
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