4.6. Aldefluor Assay

The Aldefluor kit (Stem Cell Technologies, Vancouver, BC, Canada) was used to analyse and identify cell populations with high ALDH enzymatic activity. Briefly, 1 × 10⁶ cells/mL were suspended in Aldefluor assay buffer containing the ALDH substrate BODIPY-aminoacetaldehyde and incubated at 37 °C for 45 min. For each sample, cell aliquots were incubated with or without 50 µM diethylaminobenzaldehyde (DEAB), an ALDH-specific inhibitor and 1 μM NR6. Then, the cells were analysed on (S3e Cell Sorter, BioRad, Hercules, CA, USA) and with FlowJo program v10.7.