Comparative Quantification of the Surfaceome of Human Multipotent Mesenchymal Progenitor Cells

Abstract

SummaryMesenchymal progenitor cells have great therapeutic potential, yet incomplete characterization of their cell-surface interface limits their clinical exploitation. We have employed subcellular fractionation with quantitative discovery proteomics to define the cell-surface interface proteome of human bone marrow mesenchymal stromal/stem cells (MSCs) and human umbilical cord perivascular cells (HUCPVCs). We compared cell-surface-enriched fractions from MSCs and HUCPVCs (three donors each) with adult mesenchymal fibroblasts using eight-channel isobaric-tagging mass spectrometry, yielding relative quantification on >6,000 proteins with high confidence. This approach identified 186 upregulated mesenchymal progenitor biomarkers. Validation of 10 of these markers, including ROR2, EPHA2, and PLXNA2, confirmed upregulated expression in mesenchymal progenitor populations and distinct roles in progenitor cell proliferation, migration, and differentiation. Our approach has delivered a cell-surface proteome repository that now enables improved selection and characterization of human mesenchymal progenitor populations.

Characterization of a Vimentinhigh/Nestinhigh proteome and tissue regenerative secretome generated by human pancreas-derived mesenchymal stromal cells

“…Halfon et al previously reported increased CD9 expression on the surface of skin or lung fibroblasts compared with BM‐MSCs and CD9 expression was upregulated on BM‐MSCs with increasing passage number . In contrast, Holley et al used quantitative proteomics to determine CD9 as a marker to distinguish BM‐ and adipose‐MSCs from dermal fibroblasts, although CD9 was equally enriched on perivascular cells from the umbilical cord tissue . CD146 is an established pericyte marker and robustly expressed on the surface of BM‐MSCs, which may suggest enrichment of pericytes or pericyte‐like progenitors within MSCs established from BM .…”

Myocardial regeneration strategy using Wharton’s jelly mesenchymal stem cells as an off-the-shelf ‘unlimited’ therapeutic agent: results from the Acute Myocardial Infarction First-in-Man Study

“…In short, WJMSCs are uniquely immune-privileged; they are not rejected by an allogenic host and thus their allogenic transplantation is associated with no need for immunosuppression [ 5 , 30 , 35 ]. In addition, WJMSCs can be safely expanded to “as needed” quantities [ 5 , 7 , 10 ], can be phenotypically selected on a large scale to create repositories of distinct WJMSC populations [ 36 ], and – if needed – can be further reprogrammed/engineered towards the desired cell lines [ 10 , 13 , 37 ]. Moreover, evidence is accumulating for WJMSCs’ successful use in engineering biological cardiac tissues [ 38 – 40 ] and an allogenic non-rejectable bioartificial heart [ 41 ].…”

Peer Review #2 of "A molecular classification of human mesenchymal stromal cells (v0.1)"

“…Manuscript to be reviewed forming capacity (Kaltz et al, 2010), and independently shown to be enriched more than 3 fold at protein level in bone marrow MSC compared to dermal fibroblasts or perivascular cells (Holley et al, 2015). Although several of the known and commonly used MSC markers were indeed captured in the large initial set of potential classifiers, but rejected by our statistical method on the grounds of poor selection stability, these were 'rescued' in the protein interaction network.…”

“…Other members of our network that have been previously described in human or mouse MSC biology, and used to prospectively isolate cells or have been validated at the protein level include PDGFR (Koide et al, 2007), SPINT2 (Roversi et al, 2014), CCDC80 (Charbord et al, 2015), FAP ((Bae et al, 2008), BGN (Holley et al, 2015), and TM4SF1 (Bae et al, 2011). SPINT2 is a serine protease inhibitor whose activity is required in bone-marrow MSC, and its loss alters hematopoietic stem cell function in PeerJ reviewing PDF | (2016:02:9002:1:0:REVIEW 2 Mar 2016)…”