Current Stem Cell Research & Therapy

Anthony Atala  
Wake Forest University School of Medicine,
Medical Center Boulevard
Winston Salem, NC 27157
USA

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A New Approach to CD34+ Hematopoietic Progenitor Cell Counting

Author(s): Pedro Marin, Lluis Jover, Jordi Petriz.

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

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Article Details

VOLUME: 8
ISSUE: 2
Year: 2013
Page: [163 - 171]
Pages: 9
DOI: 10.2174/1574888X11308020007
Price: $58