Clin Pediatr Hematol Oncol.  2016 Oct;23(2):125-132. 10.15264/cpho.2016.23.2.125.

Clinical Applicability of Newly Developed Image-based Cell Counter for Counting CD34+ Cells: Comparison with Flow Cytometric Analysis

• Rah WJ
• Shin EK
• Koh H
• Suh JY
• Chang M
• Nam E
• Oh JH
• Jung Y
• Lee JY
• Bong SR
• Hong SH
• Kim JY
• Han S
• Hwang JK
• Chung C
• Lee YH

• KMID: 2358368
• DOI: http://doi.org/10.15264/cpho.2016.23.2.125

Abstract

BACKGROUND
Flow cytometric analysis is the standard method for enumerating CD34+ stem cells in hematopoietic stem cell transplantation. However, it has some limitations such as expensive instrumentation, high reagent costs, and discrepancies between technicians and laboratories. We compared counts of total nucleated cells (TNCs) and CD34+ cells counts obtained from a flow cytometer with a newly-developed image-based microscopic cell counter (ADAM II) to evaluate the possibility of clinical application of the ADAM II.
METHODS
We used 18 samples of circulating peripheral blood (PB) and waste tube fractions of peripheral blood stem cells (PBSCs) harvested by apheresis after G-CSF mobilization from adult volunteer donors. We assessed the reproducibility and linearity of the new procedure and compared the numbers of TNCs and viable CD34+ cells determined with the ADAM II and two different flow cytometers (FACSCalibur, FACSCanto II).
RESULTS
Numbers of viable CD34+ cells determined with the ADAM II were accurate over the expected range; the intra-assay coefficient of variation was ≤19.8%. Linearity was also satisfactory (R²=0.99). TNC counts obtained with the ADAM II were highly correlated with those obtained with the FACSCalibur (R²>0.9841, P<0.0001) and FACSCanto II (R²>0.9620, P<0.0001), as were the numbers of viable CD34+ cells obtained with the ADAM II and the FACSCalibur and FACSCanto II (R²>0.9911, P<0.0001 and R²>0.9791, P<0.0001), respectively.
CONCLUSION
The newly developed image-based microscopic cell counter (ADAM II) appears to be suitable for enumerating TNCs and viable CD34+ cells.

Keyword

Image-based cell counter; CD34+ cell

MeSH Terms

Adult
Blood Component Removal
Cell Count*
Granulocyte Colony-Stimulating Factor
Hematopoietic Stem Cell Transplantation
Humans
Methods
Stem Cells
Tissue Donors
Volunteers
Granulocyte Colony-Stimulating Factor