I came across a sound number of techniques regarding analytical approach on cancer stem cells or cancer initiating cells (CSC). But I could not able to find out any standardized method to increase this rare but important population. using side population and CD markers only 1-20% of these cells recognized. This is very less in number for proper evaluation for therapeutic approach. Thus a technique, like use of chemical cocktail to increase this rare population by chemical induction is necessary.Kindly any one suggest a method to do enrichment of CSC in particularly in breast cancer cells.
Hi Saswat,
You can enrich your CSC population using the sphere formation assay by propagating your spheres across different generations. The more generations you have, the more enriched your CSC population is. I am attaching below two articles that might be of great interest to you.
Pastrana, E., Silva-Vargas, V., & Doetsch, F. (2011). Eyes wide open: a critical review of sphere-formation as an assay for stem cells. Cell stem cell, 8(5), 486-498.
Pattabiraman, D. R., & Weinberg, R. A. (2014). Tackling the cancer stem cells—what challenges do they pose?. Nature reviews Drug discovery, 13(7), 497-512.
Best,
Patrick
Hi Saswat,
You can enrich your CSC population using the sphere formation assay by propagating your spheres across different generations. The more generations you have, the more enriched your CSC population is. I am attaching below two articles that might be of great interest to you.
Pastrana, E., Silva-Vargas, V., & Doetsch, F. (2011). Eyes wide open: a critical review of sphere-formation as an assay for stem cells. Cell stem cell, 8(5), 486-498.
Pattabiraman, D. R., & Weinberg, R. A. (2014). Tackling the cancer stem cells—what challenges do they pose?. Nature reviews Drug discovery, 13(7), 497-512.
Best,
Patrick
1. Serial passaging in mice
Ref:https://www.researchgate.net/publication/233742776_Blebbishields_the_Emergency_Program_for_Cancer_Stem_Cells_Sphere_Formation_and_Tumorigenesis_after_Apoptosis
2. sphere formation after apoptosis (either LiCl2+Cisplatin or CHX methods)
Refs: https://www.researchgate.net/publication/233742776_Blebbishields_the_Emergency_Program_for_Cancer_Stem_Cells_Sphere_Formation_and_Tumorigenesis_after_Apoptosis
https://www.researchgate.net/publication/292615481_Mitochondrial_oligomers_boost_glycolysis_in_cancer_stem_cells_to_facilitate_blebbishield-mediated_transformation_after_apoptosis
https://www.researchgate.net/publication/299607409_Novel_PKC-z_to_p47phox_interaction_is_necessary_for_transformation_from_blebbishields
Article Blebbishields, the Emergency Program for Cancer Stem Cells: ...
Article Mitochondrial oligomers boost glycolysis in cancer stem cell...
Article Novel PKC-? to p47phox interaction is necessary for transfor...
Using the biomarker to enrich, for exmple,
Article ALDH1 is a functional marker of nasopharyngeal carcinoma ste...
Hello Saswat,
Initially growing cells in the standard serum medium and then changing the medium to one (serum free but containing growth factors like EGF, PDGF and FGF) which "selectively" allows the growth of cancer stem cells is one option that might work. Also, using ultra-low attachment dishes might help.
The attached paper might be resourceful.
Good luck!
We use the ALDH activity to FACS-sort he ALDHhigh (stem cell enriched) population in colon caner...and then we grow the sorted cells as needed or inoculate them in vivo.
Regards
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