Hello All,
I would like to know about the quickest methods as well as reagents to develop cancer stem cells from secondary cancer cell lines. Kindly attach the links for protocols if any.
Thanking you,
You may go through mammosphere formation assay. If the cell can form mammospheres it would be called to have stem cell like feature. Next, from the mammospheres, isolate single cells using cell strainers. Culture those cells and check the expression level of stem cell marker proteins such as oct4, Nanog etc.
These protocols are just one search away in google. There might be some other way also to achieve what you want. Good luck.
Hi Abdul!
The "cancer stem cell" concept is a bit nebulous so I think it would help if you defined which "stem like" characteristics you'd like to induce in your cells. Many, if not all, cancer cell lines show some "stem-like" characteristics.
Expression of genes seen in ES cells? You could use something like E-Cadherin, which is expressed on the cell surface, to purify stem like cancer cells by FACS. See available antibodies here (and at many other companies):
https://www.cellsignal.com/contents/science-cst-pathways-developmental-biology/stem-cell-and-lineage-markers/stem-cell-lineage-markers
Decrease of markers of terminal differentiation? Well... shameless plug incoming... but if you are interested in lung cancer cells, you might be able to induce such a phenotype by expression of mutant pi3k.
Article Mutationally-activated PI3’-kinase-α promotes de-differentia...
That's a pretty specific way to achieve your goal however... Might be more interesting to isolate less differentiated cells from the larger population and then figure out how they lost differentiation.
Hi Abdul- A simple exchange of growth medium with a more defined medium containing specific growth factors and maintaining the cultures for a prolonged time span can generate cancer stem cells (CSCs) in vitro. Please see below the link to the paper from our group. We have used the same technique and generated breast cancer stem cells apart from glioma CSCs. However, please note that the same medium besides generating CSCs in monolayers, also induces tumorspheres/mammospheres in culture. Please let me know if this helps. Look forward to your feedback.
Best wishes.
Article Circadian properties of cancer stem cells in glioma cell cul...
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