A secreted proteomic footprint for stem cell pluripotency
With a view to developing a much-needed non-invasive method for monitoring the healthy pluripotent state of human stem cells in culture, we undertook proteomic analysis of the waste medium from cultured embryonic (Man-13) and induced (Rebl.PAT) human pluripotent stem cells (hPSCs). Cells were grown...
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Published in | PloS one Vol. 19; no. 6; p. e0299365 |
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Main Authors | , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
Public Library of Science
14.06.2024
Public Library of Science (PLoS) |
Subjects | |
Online Access | Get full text |
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Summary: | With a view to developing a much-needed non-invasive method for monitoring the healthy pluripotent state of human stem cells in culture, we undertook proteomic analysis of the waste medium from cultured embryonic (Man-13) and induced (Rebl.PAT) human pluripotent stem cells (hPSCs). Cells were grown in E8 medium to maintain pluripotency, and then transferred to FGF2 and TGFβ deficient E6 media for 48 hours to replicate an early, undirected dissolution of pluripotency. We identified a distinct proteomic footprint associated with early loss of pluripotency in both hPSC lines, and a strong correlation with changes in the transcriptome. We demonstrate that multiplexing of four E8- against four E6- enriched secretome biomarkers provides a robust, diagnostic metric for the pluripotent state. These biomarkers were further confirmed by Western blotting which demonstrated consistent correlation with the pluripotent state across cell lines, and in response to a recovery assay. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Competing Interests: The authors have declared that no competing interests exist. Current address: Department of Psychosis Studies, Institute of Psychiatry Psychology & Neuroscience, Kings College London, London, United Kingdom Current address: Medicines Discovery Catapult, Alderley Park, Macclesfield, United Kingdom Current address: Proteomics Facility, Biomedical Sciences, University of Bristol, Bristol, United Kingdom |
ISSN: | 1932-6203 1932-6203 |
DOI: | 10.1371/journal.pone.0299365 |