Optimized Stem Cell Detection Using the DyeCycle-Triggered Side Population Phenotype

Figure 8

SP detection using the second-generation DyeCycle dyes DCG and DCO. A2780V cells harbouring an ABCB1-positive SP (a) and parental A2780 cells containing an ABCG2-positive SP (b) were stained with 500 nM of either DCG or DCO (10⁶ cells/mL) and analysed by flow cytometry using the indicated emission filters (DCO data accentuated by dotted lines). For control purpose, verapamil (A2780V) or fumitremorgin C inhibition (A2780) was performed, and drug transporters were stained with APC-conjugated monoclonal antibodies. Both DCG and DCO are able to detect ABCB1-positive SP cells, whereas neither dye can identify SP cells that express ABCG2. Note the extreme separation of ABCB1-SP cells, which made it impossible to measure the fluorescence linearly.

(a)

(b)