Standardization of a compensation matrix for the use in an ISHAGE-based single-platform for CD34+ stem cell...

Becton Dickinson (BD) FACSVia™ is a recently developed flow cytometer with which worldwide clinical experience is limited. Recently, our center started using the single-platform BD Stem Cell Kit (BD SCE) for the quantitation of viable CD34+ (7-aminoactinomycin D-7-AAD-negative cells) on a BD FACSVia™. Currently, there are no formal recommendations for the fluorescence compensation of 7-AAD in this scenario.

A 3-color fluorescence compensation matrix ("Optimized BD CS&T") was standardized based on the fluorescence of 7-AAD in samples of 27 individuals. The "Optimized BD CS&T" was compared with manual compensation (predicated method) and BD CS&T beads.

The analysis of the viable CD34+ cells/mm³ showed a very strong correlation between the manual compensation versus "Optimized BD CS&T" (r = 1.00) and manual versus BD CS&T (r = .99). The analysis of CD34+ viability (%) showed a very strong correlation for manual versus "Optimized BD CS&T" (r = .99). However, for manual versus BD CS&T, the correlation was inferior (r = .86). For viable CD34+ cells/mm³ , Bland-Altman plots showed a better concordance between manual and "Optimized BD CS&T" than between manual and BD CS&T. For CD34+ viability (%), the concordance was very good between manual and "Optimized BD CS&T", while there was a poor concordance between manual and BD CS&T.

"Optimized BD CS&T" matrix proved to be more precise than the conventional BD CS&T beads. The "Optimized BD CS&T" matrix can be used along with BD SCE kit on BD FACSVia™ flow cytometers.