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. 2011;6(5):e20399.
doi: 10.1371/journal.pone.0020399. Epub 2011 May 26.

The cell surface proteome of human mesenchymal stromal cells

Christian Niehage  1 , Charlotte Steenblock, Theresia Pursche, Martin Bornhäuser, Denis Corbeil, Bernard Hoflack
Affiliations

The cell surface proteome of human mesenchymal stromal cells

Christian Niehage et al. PLoS One. 2011.

Abstract

Background: 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.

Methodology/principal findings: 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.

Conclusions/significance: 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.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Cell surface proteome of human mesenchymal stromal cells.
Distribution and functional clustering of identified proteins.
Figure 2
Figure 2. Flow cytometric analysis of hMSC surface CD markers.
Cells were harvested and labeled with fluorescence-conjugated antibodies recognizing cell surface markers. Black: isotype control; blue: Ab against surface markers. At least three independent experiments were performed. Shown are representative flow cytometry histograms.
Figure 3
Figure 3. Comparison of hMSC and hESC cell surface protein profiles.
A. Comparison of the membrane protein profiles of hMSCs (our study) and hESCs according to . The diagram shows unique and common membrane proteins of hESCs (left) and hMSCs (right). B. Distribution and functional clustering of common membrane proteins between hESCs and hMSCs.
Figure 4
Figure 4. Differentiation of hMSCs towards the adipogenic or osteogenic lineage.
A+B. Immunocytochemistry after 1 week of differentiation. Nuclei were visualized with DAPI. C+D. Flow cytometry after 2 weeks. The change in the mean fluorescence intensity (MFI) before and after differentiation was calculated taking hMSCs as a reference. At least three independent experiments were performed. Black: hMSCs; red: differentiated cells. Shown are representative stainings.
See this image and copyright information in PMC

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