Hello all,
I'm looking to start using an ALDH assay as a marker of stem-like characteristics in Cancer Stem Cells (CSCs). These assays are quite expensive and before I commit to purchasing one, I was wondering if anyone had first hand experience with assays such as Aldefluor or Aldered. (Or any others) Do they work well? Is there an assay anyone considers to be the best or better than the rest?
Thank you for your time.
Hi Patrick,
Though I do not know what cell lines you are willing to work with, these publications might help you to get some ideas and decide upon the convenient and cost-effective option(s) that would answer your questions and serve the purpose. ICC to identify stemness by Attached-cell aldefluor method (ACAM) for ALDH1 and FACS to separate cells with high ALDH activity were carried out to identify stem-like cells in esophageal cancer cell lines. Interestingly ELISA has been used to identify ALDH1A1 (find it as a cross-reference). CD44 used as a standard stem cell marker in CSCs.
Good luck,
-Arpan
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Thanks for the reply Arpan. My boss said to go ahead and use the Stem Cell product Aldefluor as its well cited in the literature, and other products might not be accepted as readily. He wasn't as worried about the cost as I was.
Interestingly, the second paper you attached describes a process that I will be performing down the line in my experiments insofar as staining mammospheres. It'll be a nice reference. Thank you!
ALDH is linked to glycolysis shifts in CSCs and we found cells that survive apoptotic process to form spheres are CSCs (see papers from my profile). So you may try both Aldefluor assay or blebbishield-mediated sphere formation assay. You may use any drug of your choice to induce apoptosis, collect the apoptotic cells and plate it in drug free complete medium to repopulate cancer stem cells. Blebbishield-mediated sphere formation assay is cost-effective if you use cycloheximide as described in my paper below
https://www.researchgate.net/publication/292615481_Mitochondrial_oligomers_boost_glycolysis_in_cancer_stem_cells_to_facilitate_blebbishield-mediated_transformation_after_apoptosis
Article Mitochondrial oligomers boost glycolysis in cancer stem cell...
That's not the direction I'm going with my experiments but thanks for the input. Always appreciated. We've settled on Aldefluor
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