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

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...

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Autores principales: Leha, Andreas, Moens, Nathalie, Meleckyte, Ruta, Culley, Oliver J., Gervasio, Mia K., Kerz, Maximilian, Reimer, Andreas, Cain, Stuart A., Streeter, Ian, Folarin, Amos, Stegle, Oliver, Kielty, Cay M., Durbin, Richard, Watt, Fiona M., Danovi, Davide
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Academic Press 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4773406/
https://www.ncbi.nlm.nih.gov/pubmed/26608109
http://dx.doi.org/10.1016/j.ymeth.2015.11.012
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author Leha, Andreas
Moens, Nathalie
Meleckyte, Ruta
Culley, Oliver J.
Gervasio, Mia K.
Kerz, Maximilian
Reimer, Andreas
Cain, Stuart A.
Streeter, Ian
Folarin, Amos
Stegle, Oliver
Kielty, Cay M.
Durbin, Richard
Watt, Fiona M.
Danovi, Davide
author_facet Leha, Andreas
Moens, Nathalie
Meleckyte, Ruta
Culley, Oliver J.
Gervasio, Mia K.
Kerz, Maximilian
Reimer, Andreas
Cain, Stuart A.
Streeter, Ian
Folarin, Amos
Stegle, Oliver
Kielty, Cay M.
Durbin, Richard
Watt, Fiona M.
Danovi, Davide
author_sort Leha, Andreas
collection PubMed
description 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.
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spelling pubmed-47734062016-03-14 A high-content platform to characterise human induced pluripotent stem cell lines Leha, Andreas Moens, Nathalie Meleckyte, Ruta Culley, Oliver J. Gervasio, Mia K. Kerz, Maximilian Reimer, Andreas Cain, Stuart A. Streeter, Ian Folarin, Amos Stegle, Oliver Kielty, Cay M. Durbin, Richard Watt, Fiona M. Danovi, Davide Methods Article 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. Academic Press 2016-03-01 /pmc/articles/PMC4773406/ /pubmed/26608109 http://dx.doi.org/10.1016/j.ymeth.2015.11.012 Text en © 2016 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Leha, Andreas
Moens, Nathalie
Meleckyte, Ruta
Culley, Oliver J.
Gervasio, Mia K.
Kerz, Maximilian
Reimer, Andreas
Cain, Stuart A.
Streeter, Ian
Folarin, Amos
Stegle, Oliver
Kielty, Cay M.
Durbin, Richard
Watt, Fiona M.
Danovi, Davide
A high-content platform to characterise human induced pluripotent stem cell lines
title A high-content platform to characterise human induced pluripotent stem cell lines
title_full A high-content platform to characterise human induced pluripotent stem cell lines
title_fullStr A high-content platform to characterise human induced pluripotent stem cell lines
title_full_unstemmed A high-content platform to characterise human induced pluripotent stem cell lines
title_short A high-content platform to characterise human induced pluripotent stem cell lines
title_sort high-content platform to characterise human induced pluripotent stem cell lines
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4773406/
https://www.ncbi.nlm.nih.gov/pubmed/26608109
http://dx.doi.org/10.1016/j.ymeth.2015.11.012
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