Detecting human CD34+ and CD34- hematopoietic stem and progenitor cells using a Sysmex automated hematology analyzer

Area of Science:

• Hematology
• Regenerative Medicine
• Stem Cell Biology

Background:

• Accurate quantification of hematopoietic stem cells (HSC)/hematopoietic progenitor cells (HPC), including CD34+ and CD34- populations, is vital for stem cell therapies and regenerative medicine.
• Current methods for CD34+ measurement can yield differing results between automated analyzers and flow cytometry.
• Investigating novel detection methods is essential for improving the accuracy and reliability of HSC/HPC enumeration.

Purpose of the Study:

• To evaluate the utility of the immature information (IMI) channel in Sysmex XE-2100 and SE-9000 automated hematology analyzers for detecting both CD34+ and CD34- HSC/HPC.
• To compare IMI channel analysis with traditional flow cytometry for quantifying HSC/HPC populations.
• To explore the potential of automated hematology analyzers in measuring CD34-/CD133+ HSC.

Main Methods:

• Separation of CD34+ and CD34- HSC/HPC using immunologic methods.
• Quantification via flow cytometry (FACScan) and IMI channel analysis on automated hematology analyzers.
• Preparation of CD34-/CD133+ HSC via sequential antibody-based positive selection.

Main Results:

• CD34+ and CD34- HSC/HPC were successfully quantified using the IMI channel of automated hematology analyzers.
• CD34-/CD133+ HSC were observed in the same IMI channel area as CD34+ cells.
• The study confirmed the capability of automated hematology analyzers to measure both CD34+ and CD34- HSC.

Conclusions:

• Automated hematology analyzers, utilizing the IMI channel, offer a viable method for measuring both CD34+ and CD34- HSC/HPC.
• These findings may help reconcile discrepancies in HSC/HPC counts observed between automated analyzers and flow cytometry.
• Automated cell counting methods show significant potential for research and clinical applications in quantifying HSC content in cellular products.