Comparison between single‐ and dual‐platform flow cytometry for CD34 + stem cell enumeration in a pediatric population

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BACKGROUND: CD34+ hematopoietic progenitor cells (HPCs) enumeration is essential for stem cell collection adequacy. Both single (flow cytometer only) and dual-platform (flow cytometer and hematology analyzer calculation) methodologies are used, but their comparative accuracy and implications for pediatric patients remain unclear. STUDY DESIGN AND METHODS: We conducted a retrospective review of 131 autologous apheresis (HPC(A)) collections from 120 pediatric patients at a single institution (2017-2023). Assessment included differences between the two platforms as absolute and percent differences and agreement, systematic bias and association between hematology analyzers and patient characteristics with CD34+ percent differences. The impact on whether additional days of collection would be needed was assessed by comparing CD34+ dose thresholds. RESULTS: Absolute CD34+ counts were highly correlated but exhibited systematic bias. Single platform absolute CD34+ were significantly higher, especially in HPC(A). Percent differences averaged 6.3% in peripheral blood and 15.9% in HPC(A). Enumeration discrepancies varied by hematology analyzer and patient characteristics. Age, weight, and type of disease affected enumeration consistency between platforms. In 3 of 10 patients, the enumeration method potentially impacted the number of collection procedures, as the minimum CD34+ dose thresholds were met using single platform but not with dual. DISCUSSION: wEnumeration methods impact CD34+ counts and may alter clinical decisions regarding collection adequacy and need for additional days of collection. One must understand the variability in each testing methodology as it may translate to a clinically significant outcome. Method-specific adjustments or standardization protocols may be needed to ensure accurate interpretation in pediatric care.

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