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Related Experiment Videos

Single- versus dual-platform assays for human CD34+ cell enumeration.

I L Barbosa1, M E Sousa, M I Godinho

  • 1Instituto Português de Oncologia Francisco Gentil, Centro do Porto, Porto, Portugal.

Cytometry
|December 10, 1999
PubMed
Summary

Two new assays, STELLer and ProCOUNT, accurately quantify CD34+ cells. These methods show strong linear correlation and comparable performance to existing flow cytometry assays across various sample types.

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Area of Science:

  • Hematology
  • Immunology
  • Biotechnology

Background:

  • Accurate CD34+ cell quantification is crucial for stem cell transplantation and monitoring.
  • Existing methods for CD34+ cell enumeration have limitations.
  • Novel single-platform assays offer potential improvements in efficiency and accuracy.

Purpose of the Study:

  • To comparatively evaluate the performance of two new single-platform assays, IMAGN 2000 STELLer and ProCOUNT, for CD34+ cell quantification.
  • To assess the linearity, reproducibility, and correlation of these assays against a standard in-house dual-platform flow cytometric assay.
  • To analyze assay performance across different sources of CD34+ cells, including umbilical cord blood and mobilized peripheral blood.

Main Methods:

  • Comparative assessment of STELLer (microvolume fluorimetry) and ProCOUNT (flow cytometry) against an in-house dual-platform flow cytometry assay.

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  • Evaluation of assay performance using linearity tests (0-1,200 CD34+ cells/µL) and precision tests at varying concentrations.
  • Quantification of CD34+ cells in 72 samples from diverse sources: umbilical cord blood (UCB), buffy-coat (BC), mobilized peripheral blood (PB), and peripheral blood progenitor cells (PBPC).

    • Main Results:

    • All three methods demonstrated excellent linearity (R² > 0.99).
    • ProCOUNT exhibited the highest precision (CV 2.4-13.8%), followed by STELLer (3.6-26.4%) and flow cytometry (3.2-6.4%).
    • STELLer and ProCOUNT showed strong linear correlation (R > 0.90) with flow cytometry across all sample types, with no significant differences (P > 0.05), except for STELLer vs. ProCOUNT in UCB samples.

    Conclusions:

    • The STELLer and ProCOUNT assays are reliable and efficient for CD34+ cell quantification.
    • These single-platform assays demonstrate comparable performance to traditional dual-platform methods.
    • This study provides the first analysis of these assays stratified by CD34+ cell source, confirming their broad applicability.