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Graft Source

Multi-color immune-phenotyping of CD34 subsets reveals unexpected differences between various stem cell sources

Bone Marrow Transplantation volume 51, pages 1093–1100 (2016)Cite this article

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Abstract

Flow cytometric routine CD34 analysis enumerates hematopoietic stem and progenitor cells irrespective of their subpopulations although this might predict engraftment dynamics and immune reconstitution. We established a multi-color CD34 assay containing CD133, CD45RA, CD10, CD38 and CD33. We examined PBSC, donor bone marrow (BMd) and BM of patients 1 year after allografting (BM1y) regarding their CD34 subset composition, which differed significantly amongst those materials: the early CD45RA−CD133+CD38low subpopulations were significantly more frequent in PBSC than in BMd, and very low in BM1y. Vice versa, clearly more committed CD34 stages prevailed in BM, particularly in BM1y where the proportion of multi-lymphoid and CD38++ B-lymphoid precursors was highest (mean 59%). CD33 was expressed at different intensity on CD45RA±CD133± subsets allowing discrimination of earlier from more committed myeloid precursors. Compared with conventional CD34+ cell enumeration, the presented multi-color phenotyping is a qualitative approach defining different CD34 subtypes in any CD34 source. Its potential impact to predict engraftment kinetics and immune reconstitution has to be evaluated in future studies.

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Acknowledgements

We would like to thank Daniela Scharner, Dijana Trbojevic and Elke Zipperer for their excellent commitment and input when establishing the described flow cytometric assay, and for their continuous support with cell preparation and data acquisition and evaluation. Dieter Printz is particularly acknowledged for his technical support in all issues of flow cytometry.

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Authors and Affiliations

  1. Children's Cancer Research Institute, Vienna, Austria

    J Dmytrus, R Geyeregger, N Frank, C Frech & G Fritsch

  2. Department of Pediatrics, St. Anna Children's Hospital, Medical University, Vienna, Austria

    S Matthes-Martin & H Pichler

  3. Department for Blood Group Serology and Transfusion Medicine, Medical University, Vienna, Austria

    N Worel

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  1. J Dmytrus
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Correspondence to G Fritsch.

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Dmytrus, J., Matthes-Martin, S., Pichler, H. et al. Multi-color immune-phenotyping of CD34 subsets reveals unexpected differences between various stem cell sources. Bone Marrow Transplant 51, 1093–1100 (2016). https://doi.org/10.1038/bmt.2016.88

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