Coculture of equine mesenchymal stem cells and mature equine articular chondrocytes results in improved chondrogenic differentiation of the stem cells.
Abstract: Bone marrow derived mesenchymal stem cells (MSCs) can be used to repair articular cartilage defects, these cells should be properly stimulated so that they could differentiate morphologically and hold cellular synthetic features closer to maturely differentiated chondrocytes. It is well known that tissue specific environment plays an important role in cell fate determination. Once improved isolation, proliferation and differentiation protocols have been developed, the likelihood of spontaneous differentiation of MSCs into divergent lineages will be reduced, thus increasing their value for cartilage repair. The purpose of this study was to improve chondrogenic differentiation of equine MSCs using coculture with mature equine articular chondrocytes (ACs), along with the determination of the effect of adding transforming growth factor (TGF) beta1 in the pellet culture system. Following confirmation of multilineage (adipogenic, osteogenic and chondrogenic) differentiation, isolated MSCs, ACs and coculture of both cell types were transferred into pellet culture system in a DMEM-based medium supplemented with or without TGFbeta1. Chondrogenic differentiation was evaluated histologically and the relative mRNA expressions of collagen type 1 alpha1 (COL1A1), collagen type 2 alpha1(COL2A1), aggrecan (ACAN) and SRY-box 9 (SOX9) were estimated by quantitative RT-PCR. Cocultured cells showed diffuse distribution of extracellular matrix (ECM), whereas in chondrocyte pellets it was more localized to central regions. Expression of COL2A1, ACAN and SOX9 genes were higher in cocultured pellets when compared to MSCs and ACs-composed pellets. Addition of TGFbeta1 in chondrogenic differentiating medium did not consistently amplify expression of the above mentioned genes. Differentiation of equine MSCs was enhanced by coculturing in association with mature ACs, improving expression of cartilage-specific genes and producing a more homogeneous production of ECM within the newly formed cocultured cartilage. The use of the coculture system could possibly enhance the capacity of MSC-derived chondrocytes to build up stable articular cartilage-like constructs, which could play an important role in articular cartilage repair and regeneration.
Publication Date: 2010-07-22 PubMed ID: 20645581
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- Journal Article
Summary
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This research study evaluates the combined use of equine bone marrow derived mesenchymal stem cells (MSCs) and mature equine articular chondrocytes (ACs) in an effort to enhance the differentiation process of damaged articular cartilage.
Objective and Methodology
- The main aim of this study is to improve the differentiation of equine mesenchymal stem cells into articular chondrocytes through coculturing with mature chondrocytes in a pellet culture system.
- A pellet culture system incorporating mesenchymal stem cells, mature articular chondrocytes, and combination of both was used. The cells were maintained in a DMEM-based medium, optionally supplemented with transforming growth factor beta1 (TGFbeta1).
Evaluation and Results
- The differentiation process was evaluated histologically and through quantitative RT-PCR. This assessed the relative mRNA expressions corresponding to specific proteins in chondrocytes, namely collagen type 1 alpha1 (COL1A1), collagen type 2 alpha1(COL2A1), aggrecan (ACAN) and SRY-box 9 (SOX9).
- The results indicated that the cocultured cells exhibited a more homogeneous distribution of extracellular matrix (ECM), compared to chondrocyte pellets.
- The cocultured pellets also exhibited higher expression of COL2A1, ACAN, and SOX9 genes compared to the individual cell type pellets.
- However, the addition of TGFbeta1 to the differentiation medium did not consistently increase the expression levels of these genes.
Conclusions
- Coculturing equine mesenchymal stem cells with mature articular chondrocytes in a pellet culture system leads to enhanced differentiation processes, shown by increased expression of specific chondrocyte proteins and a more homogeneous production of ECM.
- The findings suggest that using a coculture system could enhance the capacity of MSC-derived chondrocytes to produce stable, articular cartilage-like constructs, and may play an important role in repairing and regenerating damaged cartilage.
Cite This Article
APA
Lettry V, Hosoya K, Takagi S, Okumura M.
(2010).
Coculture of equine mesenchymal stem cells and mature equine articular chondrocytes results in improved chondrogenic differentiation of the stem cells.
Jpn J Vet Res, 58(1), 5-15.
Publication
Researcher Affiliations
- Laboratory of Veterinary Surgery, Department of Veterinary Clinical Sciences Graduate School of Veterinary Medicine, Sapporo 060-0818, Japan.
MeSH Terms
- Aggrecans / genetics
- Animals
- Cartilage, Articular / cytology
- Cell Culture Techniques / methods
- Cell Differentiation / physiology
- Chondrocytes / cytology
- Coculture Techniques / methods
- Collagen / genetics
- DNA Primers
- Horses
- Mesenchymal Stem Cells / cytology
- Polymerase Chain Reaction
- RNA, Messenger / genetics
- SOX9 Transcription Factor / genetics
Citations
This article has been cited 18 times.- Hosseinzadeh M, Kamali A, Baghaban Eslaminejad M, Hosseini S. Higher ratios of chondrocyte to mesenchymal stem cells elevate the therapeutic effects of extracellular vesicles harvested from chondrocyte/mesenchymal stem cell co-culture on osteoarthritis in a rat model. Cell Tissue Res 2023 Aug 1;.
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