Analyses of chondrogenic induction of adipose mesenchymal stem cells by combined co-stimulation mediated by adenoviral gene transfer
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...
| Autores principales: | , , , , , , , , , |
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| Formato: | Artículo |
| Lenguaje: | inglés |
| Publicado: |
2013
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| Acceso en línea: | http://eprints.uanl.mx/14678/1/417.pdf |
| _version_ | 1824414110602756096 |
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| author | Garza Veloz, Idalia Romero Díaz, Víktor J. Martínez Fierro, Margarita de la Luz Marino Martínez, Iván Alberto González Rodríguez, Manuel Martínez Rodríguez, Herminia Guadalupe Espinoza Juárez, Marcela Alejandra Bernal Garza, Dante A. Ortiz López, Rocío Rojas Martínez, Augusto |
| author_facet | Garza Veloz, Idalia Romero Díaz, Víktor J. Martínez Fierro, Margarita de la Luz Marino Martínez, Iván Alberto González Rodríguez, Manuel Martínez Rodríguez, Herminia Guadalupe Espinoza Juárez, Marcela Alejandra Bernal Garza, Dante A. Ortiz López, Rocío Rojas Martínez, Augusto |
| author_sort | Garza Veloz, Idalia |
| collection | Repositorio Institucional |
| description | 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-b1), 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-b1, 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 cotransduced 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. |
| format | Article |
| id | eprints-14678 |
| institution | UANL |
| language | English |
| publishDate | 2013 |
| record_format | eprints |
| spelling | eprints-146782023-05-24T20:39:26Z http://eprints.uanl.mx/14678/ Analyses of chondrogenic induction of adipose mesenchymal stem cells by combined co-stimulation mediated by adenoviral gene transfer Garza Veloz, Idalia Romero Díaz, Víktor J. Martínez Fierro, Margarita de la Luz Marino Martínez, Iván Alberto González Rodríguez, Manuel Martínez Rodríguez, Herminia Guadalupe Espinoza Juárez, Marcela Alejandra Bernal Garza, Dante A. Ortiz López, Rocío Rojas Martínez, Augusto 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-b1), 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-b1, 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 cotransduced 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. 2013 Article PeerReviewed text en cc_by_nc_nd http://eprints.uanl.mx/14678/1/417.pdf http://eprints.uanl.mx/14678/1.haspreviewThumbnailVersion/417.pdf Garza Veloz, Idalia y Romero Díaz, Víktor J. y Martínez Fierro, Margarita de la Luz y Marino Martínez, Iván Alberto y González Rodríguez, Manuel y Martínez Rodríguez, Herminia Guadalupe y Espinoza Juárez, Marcela Alejandra y Bernal Garza, Dante A. y Ortiz López, Rocío y Rojas Martínez, Augusto (2013) Analyses of chondrogenic induction of adipose mesenchymal stem cells by combined co-stimulation mediated by adenoviral gene transfer. Arthritis research & therapy, 15 (4). R80. ISSN 1478-6354 http://doi.org/10.1186/ar4260 doi:10.1186/ar4260 |
| spellingShingle | Garza Veloz, Idalia Romero Díaz, Víktor J. Martínez Fierro, Margarita de la Luz Marino Martínez, Iván Alberto González Rodríguez, Manuel Martínez Rodríguez, Herminia Guadalupe Espinoza Juárez, Marcela Alejandra Bernal Garza, Dante A. Ortiz López, Rocío Rojas Martínez, Augusto Analyses of chondrogenic induction of adipose mesenchymal stem cells by combined co-stimulation mediated by adenoviral gene transfer |
| thumbnail | https://rediab.uanl.mx/themes/sandal5/images/online.png |
| title | Analyses of chondrogenic induction of adipose mesenchymal stem cells by combined co-stimulation mediated by adenoviral gene transfer |
| title_full | Analyses of chondrogenic induction of adipose mesenchymal stem cells by combined co-stimulation mediated by adenoviral gene transfer |
| title_fullStr | Analyses of chondrogenic induction of adipose mesenchymal stem cells by combined co-stimulation mediated by adenoviral gene transfer |
| title_full_unstemmed | Analyses of chondrogenic induction of adipose mesenchymal stem cells by combined co-stimulation mediated by adenoviral gene transfer |
| title_short | Analyses of chondrogenic induction of adipose mesenchymal stem cells by combined co-stimulation mediated by adenoviral gene transfer |
| title_sort | analyses of chondrogenic induction of adipose mesenchymal stem cells by combined co stimulation mediated by adenoviral gene transfer |
| url | http://eprints.uanl.mx/14678/1/417.pdf |
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