A secreted proteomic footprint for stem cell pluripotency

• Published in: PloS one Vol. 19; no. 6; p. e0299365
• Main Authors: Lewis, Philip A, Silajdžić, Edina, Smith, Helen, Bates, Nicola, Smith, Christopher A, Mancini, Fabrizio E, Knight, David, Denning, Chris, Brison, Daniel R, Kimber, Susan J
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 14.06.2024; Public Library of Science (PLoS)
• Subjects: Analysis; Automation; Biology and Life Sciences; Biomarkers; Biomarkers - metabolism; Cell culture; Cell Line; Cell lines; Computer software industry; Fibroblast growth factor 2; Fibroblast growth factors; Flow cytometry; Humans; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; Medicine and Health Sciences; Monitoring methods; Morphology; Multiplexing; Pluripotency; Pluripotent Stem Cells - cytology; Pluripotent Stem Cells - metabolism; Proteins; Proteome - analysis; Proteome - metabolism; Proteomics; Proteomics - methods; Research and Analysis Methods; Scientific equipment and supplies industry; Secretome; Stem cells; Transcriptomes; Transforming growth factors; Western blotting; United States
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0299365

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.