Has edited works of [Pierre Richer de] Belleval

Beatriz Rocha (1849771)

Cristina Ruiz-Romero (1710700)

Francisco J. Blanco (22726)

Jesús Mateos (1710703)

Patricia Fernández-Puente (1710706)

Valentina Calamia (84701)

Human mesenchymal stem cells (hMSCs), residing in bone
marrow as well as in the synovial lining of joints, can be triggered
to differentiate toward chondrocytes. Thus, hMSCs harbor great therapeutic
potential for the repair of cartilage defects in osteoarthritis (OA)
and other articular diseases. However, the molecular mechanisms underlying
the chondrogenesis process are still in part unknown. In this work,
we applied for the first time the stable isotope labeling by amino
acids in cell culture (SILAC) technique for the quantitative analysis
of protein modulation during the chondrogenic differentiation process
of hMSCs. First, we have standardized the metabolic labeling procedure
on MSCs isolated from bone marrow (hBMSCs), and we have assessed the
quality of chondrogenesis taking place in these conditions. Then,
chondrogenic differentiation was induced on these labeled cells, and
a quantitative proteomics approach has been followed to evaluate protein
changes between two differentiation days. With this strategy, we could
identify 622 different proteins by LC–MALDI-TOF/TOF analysis
and find 65 proteins whose abundance was significantly modulated between
day 2 and day 14 of chondrogenesis. Immunohistochemistry analyses
were performed to verify the changes on a panel of six proteins that
play different biological roles in the cell: fibronectin, gelsolin,
vimentin, alpha-ATPase, mitochondrial superoxide dismutase, and cyclophilin
A. All of these proteins were increased at day 14 compared to day
2 of chondrogenic induction, thus being markers of the enhanced extracellular
matrix synthesis, cell adhesion, metabolism, and response to stress
processes that take place in the early steps of chondrogenesis. Our
strategy has allowed an additional insight into both specific protein
function and the mechanisms of chondrogenesis and has provided a panel
of protein markers of this differentiation process in hBMSCs

FigShare

Metabolic Labeling of
Human Bone Marrow Mesenchymal Stem Cells for the Quantitative Analysis
of their Chondrogenic Differentiation

Broussonet, Pierre Marie Auguste

The Linnean Society of London