Chan.pdf (10.41 MB)
Multiparameter flow cytometry for the characterisation of extracellular markers on human mesenchymal stem cells
journal contribution
posted on 2014-06-16, 13:28 authored by Alexander K.C. Chan, Thomas R.J. Heathman, Karen CoopmanKaren Coopman, Christopher HewittExtracellular surface proteins are used to identify fully-functional human mesenchymal stem cells (hMSCs) in a mixed population. Here, a multiparameter flow cytometry assay was developed to examine the expression of several bone marrow-derived 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 HLA-DR were chosen as negative indicators. Single colour analysis suggested a population purity of 100 %; in contrast, when analysed via the multiparameter method, the CD73/CD105/CD90/HLA-DR/CD34 phenotypes represented 94.5 ± 1.3 % 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 pre-passage cells, highlighting the need for careful marker selection. © 2013 Springer Science+Business Media Dordrecht.
Funding
The 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).
History
School
- Aeronautical, Automotive, Chemical and Materials Engineering
Department
- Chemical Engineering
Published in
Biotechnology Letters Biotechnology LettersVolume
36Issue
4Pages
731 - 741Citation
CHAN, A.K.C. ... et al, 2014. Multiparameter flow cytometry for the characterisation of extracellular markers on human mesenchymal stem cells. Biotechnology Letters, 36 (4), pp. 731 - 741Publisher
© Springer Science+Business MediaVersion
- AM (Accepted Manuscript)
Publication date
2014Notes
The final publication is available at Springer via http://dx.doi.org/10.1007/s10529-013-1422-0ISSN
0141-5492eISSN
1573-6776Publisher version
Language
- en