Comparison of ISHAGE protocol CD34 cell enumeration with a lineage negative backgating technique

Area of Science:

• Hematopoietic Stem Cell Transplantation
• Cellular Therapy
• Immunophenotyping

Background:

• CD34(+) cell enumeration is crucial for assessing hematopoietic potential in PBSC apheresis products.
• Accurate cell counting is vital for successful transplantation outcomes.

Purpose of the Study:

• To compare the UNMC lineage negative-gating protocol with the ISHAGE protocol for CD34(+) cell enumeration.
• To evaluate the correlation of both methods with CFU-GM assay results and assess inter-technologist reproducibility.

Main Methods:

• A three-color, lineage negative-gating technique (UNMC protocol) was compared against the ISHAGE protocol.
• Over 100 apheresis products were analyzed.
• Cell doses were compared between methods and correlated with CFU-GM assay results.

Main Results:

• Both assays showed good correlation at higher CD34(+) cell doses (>0.2 x 10(6)/kg).
• The UNMC protocol demonstrated better linearity at lower doses and superior correlation with CFU-GM across all ranges.
• The UNMC protocol exhibited significantly better inter-technologist reproducibility (7% CV) compared to ISHAGE (23% CV).

Conclusions:

• The UNMC lineage negative protocol offers a larger dynamic range for CD34(+) cell enumeration.
• This protocol shows improved correlation with CFU-GM results and enhanced inter-technologist reproducibility over the ISHAGE method.