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Analyses of chondrogenic induction of adipose mesenchymal stem cells by combined co-stimulation mediated by adenoviral gene transfer

Idalia Garza-Veloz12, Viktor J Romero-Diaz3, Margarita L Martinez-Fierro2, Ivan A Marino-Martinez4, Manuel Gonzalez-Rodriguez1, Herminia G Martinez-Rodriguez1, Marcela A Espinoza-Juarez1, Dante A Bernal-Garza4, Rocio Ortiz-Lopez14 and Augusto Rojas-Martinez14*

Author Affiliations

1 Departamento de Bioquimica y Medicina Molecular, Facultad de Medicina, Universidad Autonoma de Nuevo Leon, Monterrey C.P. 64460, Mexico

2 Laboratorio de Medicina Molecular, Unidad Academica de Medicina Humana y Ciencias de la Salud, Universidad Autonoma de Zacatecas, Zacatecas, C.P 98160 Mexico

3 Departamento de Histologia, Facultad de Medicina, Universidad Autonoma de Nuevo Leon, Monterrey C.P. 64460, Mexico

4 Unidad de Terapia Genica y Celular, Centro de Investigación y Desarrollo en Ciencias de la Salud, Universidad Autonoma de Nuevo Leon, Monterrey C.P. 64460, Mexico

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Arthritis Research & Therapy 2013, 15:R80  doi:10.1186/ar4260

Published: 30 July 2013

Additional files

Additional File 1:

table describing the primers used in the qRT-PCR analysis using the iScript™ One Step RT-PCR Kit with SYBR® Green (Bio-Rad). F, forward (sense) primer; R, reverse (anti-sense) primer.

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Additional File 2:

figure showing ASC viability with single and combined adenoviral transduction. Monolayers were transduced with increasing doses of Ad.GFP, Ad.IGF-1, Ad.TGF-β1, Ad.FGF-2 and Ad.SOX9 (A) alone and (B) in combination. At 10 days, cell viability was measured with the Alamar Blue assay; the optical density OD570 to 600 nm values of untransduced cells were set as 100%. Data expressed as mean standard error of triplicate experiments. (C) At 72 hours, GFP-positive cells were counted in three fields under light and fluorescence microscopy. Results are presented as the mean percentage of fluorescent cells per field at each viral dose. (D) Representative fluorescence of ASCs transduced with 1, 10, 100, and 1,000 MOIs of Ad.GFP, as indicated.

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Additional File 3:

figure showing Protein expression from ASC aggregates after adenoviral-mediated gene transfer of IGF-1 and FGF-2 alone and in combination. Values represent levels of protein product (pg/ml) in (A,B,C) the conditioned medium at days 3, 7, 14, and 21, and (D,E,F) the aggregates at days 14 and 28. ASC aggregates singly infected with (A,D) Ad.IGF-1, (B,E) Ad.FGF-2, or (C,F) infected dually with Ad.IGF-1 at 50 MOIs and Ad.FGF-2 at 50 MOIs (100 MOIs together). Data represented as means standard deviations of three pellets per condition.

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