Abstract
Multipotent mesenchymal stromal cells (MSCs) are promising tools for regenerative medicine. They can be isolated from different sources based on their plastic-adherence property. The identification of reliable cell surface markers thus becomes the Holy Grail for their prospective isolation. Here, we determine the cell surface proteomes of human dental pulp-derived MSCs isolated from single donors after culture expansion in low (2%) or high (10%) serum-containing media. Cell surface proteins were tagged on intact cells using cell impermeable, cleavable sulfo-NHS-SS-biotin, which allows their enrichment by streptavidin pull-down. For the proteomic analyses, we first compared label-free methods to analyze cell surface proteomes i.e. composition, enrichment and proteomic differences, and we developed a new mathematical model to determine cell surface protein enrichment using a combinatorial gene ontology query. Using this workflow, we identified 101 cluster of differentiation (CD) markers and 286 non-CD cell surface proteins. Based on this proteome profiling, we identified 14 cell surface proteins, which varied consistently in abundance when cells were cultured under low or high serum conditions. Collectively, our analytical methods provide a basis for identifying the cell surface proteome of dental pulp stem cells isolated from single donors and its evolution during culture or differentiation. Our data provide a comprehensive cell surface proteome for the precise identification of dental pulp-derived MSC populations and their isolation for potential therapeutic intervention.
Highlights
Multipotent human mesenchymal stromal cells (MSCs) [1], initially described as colony-forming unit-fibroblasts [2, 3], are non-hematopoietic progenitors present in many tissues
We have previously shown that dental pulp stem cells (DPSCs) can efficiently be expanded in low serum-containing (2%) medium supplemented with epidermal growth factor (EGF) and platelet-derived growth factor BB (PDGF-BB)
We found that the best dynamic and linear range achieved by the MaxQuant intensities (MQ) intensity was covering five orders of magnitude with respect to mass input (Fig 1A and S1 Fig)
Summary
Multipotent human mesenchymal stromal cells (MSCs) [1], initially described as colony-forming unit-fibroblasts [2, 3], are non-hematopoietic progenitors present in many tissues. MSCs have the remarkable property of differentiating into a variety of cell types while self-renewing. MSCs are considered as promising candidates for tissue engineering and regenerative medicine [4], because they are able to migrate to injured tissues and to suppress responses linked with immunity [5] or inflammation [6,7,8]. Cell Surface Proteome of Dental Pulp Stem Cells study design, data collection and analysis, decision to publish, or preparation of the manuscript
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