I am already work on these markers in the isolation process of stem cells of breast cancer, and I need other markers to make isolation process perfect.
CD44highCD24low is the phenotype typically used to isolate breast cancer stem cells (BCSCs) from primary breast tumors. While it is easy for some people, it is extremely difficult for others to find this population in primary tumors. I would suggest to use a single marker Ganglioside GD2 that we recently published (Battula et al., JCI 2012) to isolate BCSCs from primary tumors and cell lines. This marker is also co-expressed on CD44highCD24low cells but it makes your life easier as it only a single marker and you dont have to worry about low or high expression as you do for CD24 and CD44. I also suggest to use CD45 a negative marker to exclude the hematopoietic cells from your sample. While staining for GD2 please use indirect staining, i.e. stain the cells first with unconjugated anti-GD2 antibody (BD) and after washing twice with PBS, stain the cells with fluorochrome conjugated secondary antibody. I found that conjugated GD2 antibodies do not work as good as unconjugated.
Article Ganglioside GD2 identifies breast cancer stem cells and prom...
CD44highCD24low is the phenotype typically used to isolate breast cancer stem cells (BCSCs) from primary breast tumors. While it is easy for some people, it is extremely difficult for others to find this population in primary tumors. I would suggest to use a single marker Ganglioside GD2 that we recently published (Battula et al., JCI 2012) to isolate BCSCs from primary tumors and cell lines. This marker is also co-expressed on CD44highCD24low cells but it makes your life easier as it only a single marker and you dont have to worry about low or high expression as you do for CD24 and CD44. I also suggest to use CD45 a negative marker to exclude the hematopoietic cells from your sample. While staining for GD2 please use indirect staining, i.e. stain the cells first with unconjugated anti-GD2 antibody (BD) and after washing twice with PBS, stain the cells with fluorochrome conjugated secondary antibody. I found that conjugated GD2 antibodies do not work as good as unconjugated.
Article Ganglioside GD2 identifies breast cancer stem cells and prom...
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