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Genet Mol Res ; 14(3): 10705-16, 2015 Sep 09.
Artículo en Inglés | MEDLINE | ID: mdl-26400300

RESUMEN

Many studies exist concerning the use of stem cells as delivery vehicles in gene therapy, expressing genes such as vascular endothelial growth factor 165 and hepatocyte growth factor. However, few reports regarding adipose tissue-derived stem cells (ADSCs) and the heme oxygenase 1 (HO-1) gene have been published. Therefore, we established a lentiviral vector encoding HO-1 and used this to infect ADSCs with the aim of producing therapeutic seed cells. In this study, ADSCs were isolated from mouse adipose tissue (AT), cultured, and identified according to the expression of antigens on their cell surface and their capacity for multilineage differentiation. A lentiviral vector encoding HO-1 was constructed, ADSCs were infected with this, and HO-1 protein expression was examined by western blotting. Our results show that ADSCs can be isolated from mouse AT, while DNA sequencing demonstrated that HO-1 was successfully transferred to the vector fused with GFP. Following 293T cell transfection, lentivirus titers were approximately 3 x 10(8) TU/mL. Fluorescence microscopy confirmed the expression of the HO-1 construct in lentivirus-infected ADSCs and the overexpression of HO-1 protein in these cells was verified by western blot. The production of ADSCs overexpressing HO-1 described in this study may aid in the development of a novel method for the treatment of asthma.


Asunto(s)
Adipocitos/metabolismo , Tejido Adiposo/metabolismo , Vectores Genéticos/química , Hemo-Oxigenasa 1/genética , Lentivirus/genética , Proteínas de la Membrana/genética , Células Madre/metabolismo , Adipocitos/citología , Tejido Adiposo/citología , Animales , Diferenciación Celular , Expresión Génica , Genes Reporteros , Vectores Genéticos/metabolismo , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Células HEK293 , Hemo-Oxigenasa 1/metabolismo , Humanos , Proteínas de la Membrana/metabolismo , Ratones , Ratones Endogámicos BALB C , Cultivo Primario de Células , Análisis de Secuencia de ADN , Células Madre/citología , Transfección
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