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Title: Multiparameter flow cytometry for the characterization of human embryonic stem cells
Authors: Brosnan, Kathryn
Want, Andrew
Coopman, Karen
Hewitt, Christopher J.
Keywords: Human embryonic stem cells
Isotype control
Multiparameter flow cytometry
SSEA 4
Tra-1-60
SSEA 1
Issue Date: 2012
Publisher: © Springer Verlag
Citation: BROSNAN, K. ... et al., 2012. Multiparameter flow cytometry for the characterization of human embryonic stem cells. Biotechnology Letters [Epub ahead of print].
Abstract: Using multiparameter staining methods and flow cytometry to investigate the pluripotency of HUES7 human embryonic stem cell cultures, it was found that the multidimensional approach of marker co-expression allowed the different cell populations to be easily identified and demonstrated cross reactivity between the SSEA 4 and SSEA 1 antibodies, resulting in a substantial false positive SSEA 1 population. It is the accepted norm to apply control gates at a 95 % confidence level of the isotype control; however, this study found that adjusting the control gate to a 99 % confidence level significantly reduced the effect of this cross reactivity. Though conversely, this gating shift also decreased the positive marker expression of SSEA 4 and Tra-1-60, indicating that there is a need for strongly expressing markers coupled with increased optimization of fluorophore/antibody combinations before a gating strategy of 99 % can be implemented on a more routine basis.
Description: This article was accepted for publication in the journal, Biotechnology Letters [© Springer Verlag]. The journal's website is at: http://www.springerlink.com/content/0141-5492/
Version: Accepted for publication
URI: https://dspace.lboro.ac.uk/2134/11056
ISSN: 0141-5492
Appears in Collections:Published Articles (Chemical Engineering)

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