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Title: Analyses of chondrogenic induction of adipose mesenchymal stem cells by combined costimulation mediated by adenoviral gene transfer
Authors: Garza Veloz, Idalia
Romero Díaz, Viktor J.
Martínez Fierro, Margarita de la Luz
Marino Martínez, Iván
González Rodríguez, Manuel
Martínez Rodríguez, Herminia Guadalupe
Espinoza Juárez, Marcela
Bernal Garza, Dante
Ortíz López, Rocío
Rojas Martínez, Augusto
Issue Date: 30-Jul-2013
Publisher: Springer
Abstract: 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 pointfor 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 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.
ISSN: 1478-6362
Other Identifiers: info:eu-repo/semantics/publishedVersion
Appears in Collections:*Documentos Académicos*-- Doc. en Ing. y Tec. Aplicada

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