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Comparison of Simultaneous CD34+ and CD3+ Quantification with a Modified Stem Cell Enumeration Kit on Two Different
Clinical Laboratory
|February 7, 2017
Summary
A modified ISHAGE protocol accurately quantifies CD3+ and CD34+ cells in peripheral blood stem cell apheresis using two flow cytometers. Bead quantification ensures reliable results across both machines, crucial for donor lymphocyte infusions.
Area of Science:
- Hematology
- Immunology
- Flow Cytometry
Background:
- Peripheral blood stem cell apheresis requires CD34+ cell quantification via flow cytometry (ISHAGE protocol).
- Peripheral blood stem cell concentrates (PBSC) contain T cells, valuable for donor lymphocyte infusions.
- Quantifying both CD3+ and CD34+ cells in allogeneic PBSC is clinically relevant.
Purpose of the Study:
- To evaluate a modified ISHAGE protocol for simultaneous CD3+ and CD34+ cell quantification.
- To compare the performance of the modified protocol on FACSCalibur and FACSVerse flow cytometers.
Main Methods:
- Utilized standard and modified ISHAGE protocols with CD3+ cell quantification.
- Employed bead-based enumeration kits (Trucount) for cell concentration measurements.
- Compared bead-based quantification with a volume sensor on the FACSVerse.
Main Results:
- No significant differences in cell counts between standard and modified ISHAGE protocols using bead quantification.
- Consistent results were observed between FACSCalibur and FACSVerse when using the modified protocol.
- Significant differences were found between bead and volume sensor quantification on the FACSVerse.
Conclusions:
- The modified ISHAGE protocol is effective for CD3+ and CD34+ cell quantification on both tested flow cytometers.
- Bead-based quantification is recommended for reliable and consistent results with the modified protocol.
