During preliminary exposure of these cells to FGF, EGF, B27, and insulin factors; only a small amount of cancer stem cells was detected by flow cytometry analysis (CD133, CD44 positive cells) after two weeks of treatment. Meanwhile, I need a large number of cancer stem cells (over than 10%) to enrich them by MACS procedure?
You are right. In a cell line the cancer stem cells are observed as small fractions because the self-renewal divisions constantly produce differentiated or bulk cancer cells in addition to cancer stem cells. Again it depends on cell type and the marker you choose to isolate them. Once you isolate them you may try to maintain stemness by appropriate treatments/culture conditions that are specific for the cancer stem cells. The catch is that those maintenance characteristics are poorly understood for many cancer stem cell types. Some cells respond to pH (despite the controversy of STAP cells), some cells respond to growth factors or intracellular signaling.
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