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Title of Journal: Biotechnol Lett

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Abbravation: Biotechnology Letters

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Springer Netherlands

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DOI

10.1007/s10444-013-9325-0

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1573-6776

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Multiparameter flow cytometry for the characterisa

Authors: Alexander K C Chan Thomas R J Heathman Karen Coopman Christopher J Hewitt
Publish Date: 2013/12/10
Volume: 36, Issue: 4, Pages: 731-741
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Abstract

Extracellular surface proteins are used to identify fullyfunctional human mesenchymal stem cells hMSCs in a mixed population Here a multiparameter flow cytometry assay was developed to examine the expression of several bone marrowderived hMSC markers simultaneously at the single cell level The multiparameter approach demonstrates a depth of analysis that goes far beyond the conventional single or dual staining methods CD73 CD90 and CD105 were chosen as positive markers as they are expressed on multipotent hMSCs whilst CD34 and HLADR were chosen as negative indicators Single colour analysis suggested a population purity of 100  in contrast when analysed via the multiparameter method the CD73+ve/CD105+ve/CD90+ve/HLADR−ve/CD34−ve phenotypes represented 945 ± 13  of the total cell population Also although CD271 has been posited as a definite early stage hMSC marker here we show it is not present on prepassage cells highlighting the need for careful marker selectionThe authors acknowledge funding support from the Engineering and Physical Sciences Research Council EPSRC Doctoral Training Centre in Regenerative Medicine and Bioprocessing Research Industries Club BRIC The authors would also like to thank Andrew Want Merck Millipore UK for his training and input to the study design

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