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Evaluation of the BD Stem Cell Enumeration Kit on the BD FACSCanto II flow cytometer using bd facscanto clinical and
Vladimira Rimac1, Ines Bojanić1, Koraljka Gojčeta1
1Clinical Department of Transfusion Medicine and Transplantation Biology, University Hospital Centre Zagreb, Zagreb, Croatia.
Introduction:
CD34+ hematopoietic stem cell (HSC) enumeration by cell flow cytometry is routinely used in clinical laboratories for monitoring of HSC mobilization into peripheral blood and assessment of the quality of HSC products. The modified ISHAGE protocol is the most often used procedure for determination of CD34+ cells using flow cytometry. The aim of this study was to evaluate BD Enumeration stem cell kit on flow cytometer BD facscanto II, using facscanto clinical and facsdiva softwares.
Methods:
Validation study included determination of within-run and between-run precision, trueness (bias), comparison of the test results analyzed on facscanto clinical and facsdiva softwares, assessment of linearity, specimen stability, and carryover.
Results:
For between-run precision, coefficients of variation (CVs) were all <10%, except for low control level on facsdiva software. CVs for within-run precision were <10%, except for high absolute count of CD34+ cells on facsdiva software. Comparison of data showed no statistically significant differences between facscanto clinical and facsdiva software (Spearman's rank correlation coefficients were .993 for % of CD34+ cells and 0.983 for absolute count of CD34+ cells). In linearity study, bias for all dilutions was < 20%, and carryover assessment cannot be considered significant on both softwares. There was a statistically significant difference (P = .044) in absolute count of CD34+ cells after 24 hours of storage, when using facscanto clinical software.
Conclusion:
BD Stem Cell Enumeration Kit can be used in routine laboratory work on BD FACSCanto II instrument, whereas facscanto clinical and facsdiva software were used for acquisition and data analysis.
