Journal of Oral Tissue Engineering

ORIGINAL ARTICLE
Highly Expression and Biological Function of Type VI Collagen on the Early Events of Chondrogenesis in Human Mesenchymal Stem Cells

Jiefeng CUI1, 2, Yinkun LIU2, Rena MATUMOTO1 and Toshimasa UEMURA1
1Nanosystem Research Institute (NRI), National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan
2Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai, China


J Oral Tissue Engin 2013;11(1):29-41

SYNOPSIS
Previously we reported a successful three-dimensional cartilage regeneration using an RWV (rotation wall vessel) bioreactor from human mesenchymal stem cells (hMSCs)(Sakai et al. J Orthop Res. 27, 2009), which is useful for better un-derstanding the initiation mechanism of chondrogenic differentiation of hMSCs and discovering its associated proteins. Entire proteome of hMSCs and spherical car-tilaginous constructs by RWV culture was comparatively analyzed using two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption ionization/ time-of-flight/mass spectrometry (MALDI-TOF-MS/MS) to determine candidate proteins associated with early chondrogenesis. Among them, there found a greatly up-regulation of COL6A1 and COL6A2 proteins on day 10, which encode matrix proteins with great changes at early stage of chondrogenesis in parallel with the increase of SOX9. A specific antibody against to the extracellular protein COL6 effectively postponed the progress of early chondrogenesis, fol-lowed by the successive reduction of extracellular COL6 in spite of a temporary increase in genes expression and up-regulation of SOX9 phosphorylation on the early stage of antibody neutralization. It suggested that the extracellular matrix protein COL6 might be a potential key molecule to involve in early events of chondrogenesis via regulating or influencing the expression of SOX9.

Key words: MSCs, chondrogenesis, Type VI collagen, comparative proteomics