Analyses of chondrogenic induction of adipose mesenchymal stem cells by combined co-stimulation mediated by adenoviral gene transfer
| Autor: | Idalia Garza-Veloz, Manuel Gonzalez-Rodriguez, Margarita L Martinez-Fierro, Iván A. Marino-Martínez, Marcela A. Espinoza-Juarez, Viktor J Romero-Diaz, Augusto Rojas-Martinez, Rocio Ortiz-Lopez, Dante A Bernal-Garza, Herminia G. Martínez-Rodríguez |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
Cell Survival
Cellular differentiation Blotting Western Immunology Real-Time Polymerase Chain Reaction Adenoviridae Transforming Growth Factor beta1 Chondrocytes Rheumatology Transduction Genetic medicine Animals Humans Immunology and Allergy Insulin-Like Growth Factor I Fibroblast Cells Cultured Aggrecan Sheep biology Biglycan Cartilage Mesenchymal stem cell Gene Transfer Techniques Cell Differentiation Mesenchymal Stem Cells SOX9 Transcription Factor Chondrogenesis Immunohistochemistry Molecular biology medicine.anatomical_structure Adipose Tissue Proteoglycan biology.protein Fibroblast Growth Factor 2 Research Article |
| Zdroj: | Arthritis Research & Therapy |
| ISSN: | 1478-6354 |
| DOI: | 10.1186/ar4260 |
| Popis: | Introduction Adipose-derived stem cells (ASCs) have the potential to differentiate into cartilage under stimulation with some reported growth and transcriptional factors, which may constitute an alternative for cartilage replacement approaches. In this study, we analyzed the in vitro chondrogenesis of ASCs transduced with adenoviral vectors encoding insulin-like growth factor-1 (IGF-1), transforming growth factor beta-1 (TGF-β1), fibroblast growth factor-2 (FGF-2), and sex-determining region Y-box 9 (SOX9) either alone or in combinations. Methods Aggregate cultures of characterized ovine ASCs were transduced with 100 multiplicity of infections of Ad.IGF-1, Ad.TGF-β1, Ad.FGF-2, and Ad.SOX9 alone or in combination. These were harvested at various time points for detection of cartilage-specific genes expression by quantitative real-time PCR or after 14 and 28 days for histologic and biochemical analyses detecting proteoglycans, collagens (II, I and X), and total sulfated glycosaminoglycan and collagen content, respectively. Results Expression analyses showed that co-expression of IGF-1 and FGF-2 resulted in higher significant expression levels of aggrecan, biglycan, cartilage matrix, proteoglycan, and collagen II (all P ≤0.001 at 28 days). Aggregates co-transduced with Ad.IGF-1/Ad.FGF-2 showed a selective expression of proteoglycans and collagen II, with limited expression of collagens I and × demonstrated by histological analyses, and had significantly greater glycosaminoglycan and collagen production than the positive control (P ≤0.001). Western blot analyses for this combination also demonstrated increased expression of collagen II, while expression of collagens I and × was undetectable and limited, respectively. Conclusion Combined overexpression of IGF-1/FGF-2 within ASCs enhances their chondrogenic differentiation inducing the expression of chondrogenic markers, suggesting that this combination is more beneficial than the other factors tested for the development of cell-based therapies for cartilage repair. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink