title: Microenvironmental changes during differentiation of mesenchymal stem cells towards chondrocytes.
abstract: Chondrogenesis is a process involving stem-cell differentiation through the coordinated effects of growth/differentiation factors and extracellular matrix (ECM) components. Recently, mesenchymal stem cells (MSCs) were found within the cartilage, which constitutes a specific niche composed of ECM proteins with unique features. Therefore, we hypothesized that the induction of MSC differentiation towards chondrocytes might be induced and/or influenced by molecules from the microenvironment. Using microarray analysis, we previously identified genes that are regulated during MSC differentiation towards chondrocytes. In this study, we wanted to precisely assess the differential expression of genes associated with the microenvironment using a large-scale real-time PCR assay, according to the simultaneous detection of up to 384 mRNAs in one sample. Chondrogenesis of bone-marrow-derived human MSCs was induced by culture in micropellet for various periods of time. Total RNA was extracted and submitted to quantitative RT-PCR. We identified molecules already known to be involved in attachment and cell migration, including syndecans, glypicans, gelsolin, decorin, fibronectin, and type II, IX and XI collagens. Importantly, we detected the expression of molecules that were not previously associated with MSCs or chondrocytes, namely metalloproteases (MMP-7 and MMP-28), molecules of the connective tissue growth factor (CTGF); cef10/cyr61 and nov (CCN) family (CCN3 and CCN4), chemokines and their receptors chemokine CXC motif ligand (CXCL1), Fms-related tyrosine kinase 3 ligand (FlT3L), chemokine CC motif receptor (CCR3 and CCR4), molecules with A Disintegrin And Metalloproteinase domain (ADAM8, ADAM9, ADAM19, ADAM23, A Disintegrin And Metalloproteinase with thrombospondin type 1 motif ADAMTS-4 and ADAMTS-5), cadherins (4 and 13) and integrins (alpha4, alpha7 and beta5). Our data suggest that crosstalk between ECM components of the microenvironment and MSCs within the cartilage is responsible for the differentiation of MSCs into chondrocytes.
authors: Inserm, U 844, 80 avenue Augustin Fliche, Montpellier, F-34091 France. djouad@montp.inserm.fr
authors: Farida
authors: F
authors: Djouad
authors: Bruno
authors: B
authors: Delorme
authors: Marielle
authors: M
authors: Maurice
authors: Claire
authors: C
authors: Bony
authors: Florence
authors: F
authors: Apparailly
authors: Pascale
authors: P
authors: Louis-Plence
authors: François
authors: F
authors: Canovas
authors: Pierre
authors: P
authors: Charbord
authors: Danièle
authors: D
authors: Noël
authors: Christian
authors: C
authors: Jorgensen
mesh_terms: D000328:Adult; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis; D002356:Cartilage / Q000166:cytology / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000096:biosynthesis; D002454:Cell Differentiation; D018925:Chemokines / Q000096:biosynthesis; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D005434:Flow Cytometry; D015870:Gene Expression; D020869:Gene Expression Profiling / Q000379:methods; D006801:Humans; D016023:Integrins / Q000096:biosynthesis; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis*; D019707:Receptors, Chemokine / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D016212:Transforming Growth Factor beta / Q000096:biosynthesis
chemical_list: C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D015815:Cell Adhesion Molecules; D018925:Chemokines; D016023:Integrins; D012333:RNA, Messenger; D019707:Receptors, Chemokine; D016212:Transforming Growth Factor beta