Abstract
Background: In the absence of a gold standard for hematopoietic progenitor counting, the intra-laboratory variation between commonly used strategies for progenitor assessment was compared. Methods and Materials: We used a pool of FITC-conjugated monoclonal antibodies (CD14, CD11b) and PE-CD34 to facilitate CD34 counting, excluding CD14+/CD11b+ bright cells. We compared this protocol with other common methodologies, such as the single-staining approach, known as the Milan method, and the ISHAGE multiparameter method. Results: We show that the CD14/CD11b protocol is a valid approach to progenitor cell counting. Though different methods give different results for progenitor cell counting, Lin’s coefficient shows high concordance among flow cytometry counts but a substantial difference with colony- forming unit counts. Both the ISHAGE and CD14/CD11b protocol give higher counts than the Milan method. Moreover, on revising the ISHAGE analysis, we described a rare population of cells with the CD34+CD45neg phenotype, which could have an impact in CD34 counting. Conclusions: We have designed a valid alternative approach for hematopoietic progenitor cell counting, and we show that different methods give different results.
Keywords: CD34, CD45, flow cytometry, ISHAGE, Milan, quality control, Hematopoietic Progenitor Cell Counting, monoclonal antibodies
Current Stem Cell Research & Therapy
Title:A New Approach to CD34+ Hematopoietic Progenitor Cell Counting
Volume: 8 Issue: 2
Author(s): Pedro Marin, Lluis Jover and Jordi Petriz
Affiliation:
Keywords: CD34, CD45, flow cytometry, ISHAGE, Milan, quality control, Hematopoietic Progenitor Cell Counting, monoclonal antibodies
Abstract: Background: In the absence of a gold standard for hematopoietic progenitor counting, the intra-laboratory variation between commonly used strategies for progenitor assessment was compared. Methods and Materials: We used a pool of FITC-conjugated monoclonal antibodies (CD14, CD11b) and PE-CD34 to facilitate CD34 counting, excluding CD14+/CD11b+ bright cells. We compared this protocol with other common methodologies, such as the single-staining approach, known as the Milan method, and the ISHAGE multiparameter method. Results: We show that the CD14/CD11b protocol is a valid approach to progenitor cell counting. Though different methods give different results for progenitor cell counting, Lin’s coefficient shows high concordance among flow cytometry counts but a substantial difference with colony- forming unit counts. Both the ISHAGE and CD14/CD11b protocol give higher counts than the Milan method. Moreover, on revising the ISHAGE analysis, we described a rare population of cells with the CD34+CD45neg phenotype, which could have an impact in CD34 counting. Conclusions: We have designed a valid alternative approach for hematopoietic progenitor cell counting, and we show that different methods give different results.
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Cite this article as:
Marin Pedro, Jover Lluis and Petriz Jordi, A New Approach to CD34+ Hematopoietic Progenitor Cell Counting, Current Stem Cell Research & Therapy 2013; 8 (2) . https://dx.doi.org/10.2174/1574888X11308020007
DOI https://dx.doi.org/10.2174/1574888X11308020007 |
Print ISSN 1574-888X |
Publisher Name Bentham Science Publisher |
Online ISSN 2212-3946 |
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