A secreted proteomic footprint for stem cell pluripotency

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

doi:10.1371/journal.pone.0299365

A secreted proteomic footprint for stem cell pluripotency

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^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

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.

Original language	English
Journal	PLoS ONE
Volume	19
Issue number	6
Early online date	14 Jun 2024
DOIs	https://doi.org/10.1371/journal.pone.0299365
Publication status	E-pub ahead of print - 14 Jun 2024

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title = "A secreted proteomic footprint for stem cell pluripotency",

abstract = "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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T1 - A secreted proteomic footprint for stem cell pluripotency

AU - Lewis, Philip A.

AU - Silajdžić, Edina

AU - Smith, Helen

AU - Bates, Nicola

AU - Smith, Christopher A.

AU - Mancini, Fabrizio E.

AU - Knight, David

AU - Denning, Chris

AU - Brison, Daniel R.

AU - Kimber, Susan J.

PY - 2024/6/14

Y1 - 2024/6/14

N2 - 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.

AB - 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.

U2 - 10.1371/journal.pone.0299365

DO - 10.1371/journal.pone.0299365

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SN - 1932-6203

VL - 19

JO - PLoS ONE

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A secreted proteomic footprint for stem cell pluripotency

Abstract

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