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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| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2024-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0299365 |
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| author | Philip A Lewis Edina Silajdžić Helen Smith Nicola Bates Christopher A Smith Fabrizio E Mancini David Knight Chris Denning Daniel R Brison Susan J Kimber |
| author_facet | Philip A Lewis Edina Silajdžić Helen Smith Nicola Bates Christopher A Smith Fabrizio E Mancini David Knight Chris Denning Daniel R Brison Susan J Kimber |
| author_sort | Philip A Lewis |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-ab5be9b6d876451482c5833fcfe03b9d |
| institution | Kabale University |
| issn | 1932-6203 |
| language | English |
| publishDate | 2024-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-ab5be9b6d876451482c5833fcfe03b9d2025-01-04T05:31:02ZengPublic Library of Science (PLoS)PLoS ONE1932-62032024-01-01196e029936510.1371/journal.pone.0299365A secreted proteomic footprint for stem cell pluripotency.Philip A LewisEdina SilajdžićHelen SmithNicola BatesChristopher A SmithFabrizio E ManciniDavid KnightChris DenningDaniel R BrisonSusan J KimberWith 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.https://doi.org/10.1371/journal.pone.0299365 |
| spellingShingle | Philip A Lewis Edina Silajdžić Helen Smith Nicola Bates Christopher A Smith Fabrizio E Mancini David Knight Chris Denning Daniel R Brison Susan J Kimber A secreted proteomic footprint for stem cell pluripotency. PLoS ONE |
| title | A secreted proteomic footprint for stem cell pluripotency. |
| title_full | A secreted proteomic footprint for stem cell pluripotency. |
| title_fullStr | A secreted proteomic footprint for stem cell pluripotency. |
| title_full_unstemmed | A secreted proteomic footprint for stem cell pluripotency. |
| title_short | A secreted proteomic footprint for stem cell pluripotency. |
| title_sort | secreted proteomic footprint for stem cell pluripotency |
| url | https://doi.org/10.1371/journal.pone.0299365 |
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