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Methods. 2016 Mar 1;96:85-96. doi: 10.1016/j.ymeth.2015.11.012. Epub 2015 Nov 25.

A high-content platform to characterise human induced pluripotent stem cell lines.

Author information

1
Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SD, UK.
2
HipSci Cell Phenotyping, Centre for Stem Cells and Regenerative Medicine, King's College London, Great Maze Pond, London SE1 9RT, UK.
3
HipSci Cell Phenotyping, Centre for Stem Cells and Regenerative Medicine, King's College London, Great Maze Pond, London SE1 9RT, UK; NIHR Biomedical Research Centre for Mental Health Informatics Core, King's College London, De Crespigny Park, London SE5 8AF, UK.
4
Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, United Kingdom.
5
European Molecular Biology Laboratory, European Bioinformatics Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SA, UK.
6
NIHR Biomedical Research Centre for Mental Health Informatics Core, King's College London, De Crespigny Park, London SE5 8AF, UK.
7
HipSci Cell Phenotyping, Centre for Stem Cells and Regenerative Medicine, King's College London, Great Maze Pond, London SE1 9RT, UK. Electronic address: davide.danovi@kcl.ac.uk.

Abstract

Induced pluripotent stem cells (iPSCs) provide invaluable opportunities for future cell therapies as well as for studying human development, modelling diseases and discovering therapeutics. In order to realise the potential of iPSCs, it is crucial to comprehensively characterise cells generated from large cohorts of healthy and diseased individuals. The human iPSC initiative (HipSci) is assessing a large panel of cell lines to define cell phenotypes, dissect inter- and intra-line and donor variability and identify its key determinant components. Here we report the establishment of a high-content platform for phenotypic analysis of human iPSC lines. In the described assay, cells are dissociated and seeded as single cells onto 96-well plates coated with fibronectin at three different concentrations. This method allows assessment of cell number, proliferation, morphology and intercellular adhesion. Altogether, our strategy delivers robust quantification of phenotypic diversity within complex cell populations facilitating future identification of the genetic, biological and technical determinants of variance. Approaches such as the one described can be used to benchmark iPSCs from multiple donors and create novel platforms that can readily be tailored for disease modelling and drug discovery.

KEYWORDS:

Cell based assays; High content; Human pluripotent stem cells; Induced pluripotent stem cells; Phenotype screening; iPSCs

PMID:
26608109
PMCID:
PMC4773406
DOI:
10.1016/j.ymeth.2015.11.012
[Indexed for MEDLINE]
Free PMC Article

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