The cell surface proteome of human mesenchymal stromal cells

• Published in: PloS one 2011-05, Vol.6 (5), p.e20399-e20399
• Main Authors: Niehage, Christian, Steenblock, Charlotte, Pursche, Theresia, Bornhäuser, Martin, Corbeil, Denis, Hoflack, Bernard
• Format: Article
• Language: English
• Subjects: Adipogenesis; Adult; Analysis; Analytical methods; Antibodies; Antigens, CD - metabolism; Arthritis; Biocompatibility; Biological products; Biology; Biomarkers; Biomarkers - metabolism; Biomedical materials; Biotechnology; Biotin; Biotinylation; Bone marrow; Cardiomyocytes; Cell adhesion; Cell adhesion & migration; Cell adhesion molecules; Cell Lineage; Cell Membrane - metabolism; Cell surface; Datasets; Embryo cells; Embryonic stem cells; Embryonic Stem Cells - metabolism; Epidermal growth factor; Flow Cytometry; Gene expression; Humans; Ligands; Mass spectrometry; Mass spectroscopy; Membrane proteins; Membrane Proteins - isolation & purification; Membrane Proteins - metabolism; Mesenchymal stem cells; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - metabolism; Mesenchyme; Motility; Neutrophils; Osteogenesis; Plasma; Plastics; Populations; Proteins; Proteome - isolation & purification; Proteome - metabolism; Proteomes; Receptors; Regenerative medicine; Signaling; Stem cells; Stromal cells; Stromal Cells - cytology; Stromal Cells - metabolism; Surface markers; Tissue engineering; Transcription factors
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0020399

Multipotent human mesenchymal stromal cells (hMSCs) are considered as promising biological tools for regenerative medicine. Their antibody-based isolation relies on the identification of reliable cell surface markers. To obtain a comprehensive view of the cell surface proteome of bone marrow-derived hMSCs, we have developed an analytical pipeline relying on cell surface biotinylation of intact cells using cell impermeable, cleavable sulfo-NHS-SS-biotin to enrich the plasma membrane proteins and mass spectrometry for identification with extremely high confidence. Among the 888 proteins identified, we found ≈200 bona fide plasma membrane proteins including 33 cell adhesion molecules and 26 signaling receptors. In total 41 CD markers including 5 novel ones (CD97, CD112, CD239, CD276, and CD316) were identified. The CD markers are distributed homogenously within plastic-adherent hMSC populations and their expression is modulated during the process of adipogenesis or osteogenesis. Moreover, our in silico analysis revealed a significant difference between the cell surface proteome of hMSCs and that of human embryonic stem cells reported previously. Collectively, our analytical methods not only provide a basis for further studies of mechanisms maintaining the multipotency of hMSCs within their niches and triggering their differentiation after signaling, but also a toolbox for a refined antibody-based identification of hMSC populations from different tissues and their isolation for therapeutic intervention.