title: Multiplex High-Throughput Targeted Proteomic Assay To Identify Induced Pluripotent Stem Cells.
abstract: Induced pluripotent stem cells have great potential as a human model system in regenerative medicine, disease modeling, and drug screening. However, their use in medical research is hampered by laborious reprogramming procedures that yield low numbers of induced pluripotent stem cells. For further applications in research, only the best, competent clones should be used. The standard assays for pluripotency are based on genomic approaches, which take up to 1 week to perform and incur significant cost. Therefore, there is a need for a rapid and cost-effective assay able to distinguish between pluripotent and nonpluripotent cells. Here, we describe a novel multiplexed, high-throughput, and sensitive peptide-based multiple reaction monitoring mass spectrometry assay, allowing for the identification and absolute quantitation of multiple core transcription factors and pluripotency markers. This assay provides simpler and high-throughput classification into either pluripotent or nonpluripotent cells in 7 min analysis while being more cost-effective than conventional genomic tests.
authors.affiliation: Centre for Translational Omics, UCL Great Ormond Street Institute of Child Health , London, WC1N 1EH, United Kingdom.
authors.forename: Anna
authors.initials: A
authors.lastname: Baud
authors.affiliation: Department of Physiology, Anatomy & Genetics, Oxford University , Oxford, OX1 3PT, United Kingdom.
authors.forename: Frank
authors.initials: F
authors.lastname: Wessely
authors.affiliation: Centre for Translational Omics, UCL Great Ormond Street Institute of Child Health , London, WC1N 1EH, United Kingdom.
authors.forename: Francesca
authors.initials: F
authors.lastname: Mazzacuva
authors.affiliation: Centre for Translational Omics, UCL Great Ormond Street Institute of Child Health , London, WC1N 1EH, United Kingdom.
authors.forename: James
authors.initials: J
authors.lastname: McCormick
authors.affiliation: Centre for Translational Omics, UCL Great Ormond Street Institute of Child Health , London, WC1N 1EH, United Kingdom.
authors.forename: Stephane
authors.initials: S
authors.lastname: Camuzeaux
authors.affiliation: Centre for Translational Omics, UCL Great Ormond Street Institute of Child Health , London, WC1N 1EH, United Kingdom.
authors.forename: Wendy E
authors.initials: WE
authors.lastname: Heywood
authors.affiliation: MRC Laboratory for Molecular Cell Biology, University College London , London, WC1E 6BT, United Kingdom.
authors.forename: Daniel
authors.initials: D
authors.lastname: Little
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford , Oxford, OX1 3QX, United Kingdom.|James Martin Stem Cell Facility, Sir William Dunn School of Pathology, University of Oxford , Oxford, OX1 3RE, United Kingdom.
authors.forename: Jane
authors.initials: J
authors.lastname: Vowles
authors.affiliation: Janssen Research and Development , Beerse, 2340, Belgium.
authors.forename: Marianne
authors.initials: M
authors.lastname: Tuefferd
authors.affiliation: MRC Laboratory for Molecular Cell Biology, University College London , London, WC1E 6BT, United Kingdom.
authors.forename: Olukunbi
authors.initials: O
authors.lastname: Mosaku
authors.affiliation: Institute of Genetic Medicine, Newcastle University , Newcastle, NE1 3BZ, United Kingdom.
authors.forename: Majlinda
authors.initials: M
authors.lastname: Lako
authors.affiliation: Institute of Genetic Medicine, Newcastle University , Newcastle, NE1 3BZ, United Kingdom.
authors.forename: Lyle
authors.initials: L
authors.lastname: Armstrong
authors.affiliation: Department of Physiology, Anatomy & Genetics, Oxford University , Oxford, OX1 3PT, United Kingdom.
authors.forename: Caleb
authors.initials: C
authors.lastname: Webber
authors.affiliation: The Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital , Oxford, OX3 9DS, United Kingdom.
authors.forename: M Zameel
authors.initials: MZ
authors.lastname: Cader
authors.affiliation: Janssen Research and Development , Beerse, 2340, Belgium.
authors.forename: Pieter
authors.initials: P
authors.lastname: Peeters
authors.affiliation: MRC Laboratory for Molecular Cell Biology, University College London , London, WC1E 6BT, United Kingdom.
authors.forename: Paul
authors.initials: P
authors.lastname: Gissen
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford , Oxford, OX1 3QX, United Kingdom.|James Martin Stem Cell Facility, Sir William Dunn School of Pathology, University of Oxford , Oxford, OX1 3RE, United Kingdom.
authors.forename: Sally A
authors.initials: SA
authors.lastname: Cowley
authors.affiliation: Centre for Translational Omics, UCL Great Ormond Street Institute of Child Health , London, WC1N 1EH, United Kingdom.
authors.forename: Kevin
authors.identifier: 0000-0003-0763-8288
authors.initials: K
authors.lastname: Mills
mesh_terms: D002454:Cell Differentiation; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods; D020543:Proteome / Q000032:analysis* / Q000378:metabolism; D040901:Proteomics; D012867:Skin / Q000166:cytology; D014157:Transcription Factors / Q000032:analysis / Q000378:metabolism
chemical_list: D020543:Proteome; D014157:Transcription Factors