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  • Title: Lentiviral-mediated gene transfer into human adipose-derived stem cells: role of NELL1 versus BMP2 in osteogenesis and adipogenesis in vitro.
    Author: Liu Y, Chen C, He H, Wang D, E L, Liu Z, Liu H.
    Journal: Acta Biochim Biophys Sin (Shanghai); 2012 Oct; 44(10):856-65. PubMed ID: 23017834.
    Abstract:
    NEL-like molecule 1 (NELL1) is a potent osteogenic factor associated with craniosynostosis. Adenoviruses, the most commonly used viral vectors for gene therapy, have several disadvantages that may restrict osteogenesis. Previous studies have shown that lentiviruses can serve as ideal vectors for gene therapy for bone regeneration. In this study, two lentiviral vectors (LvNELL1 and LvBMP2) that encode human NELL1 and bone morphogenetic protein-2 (BMP2), respectively, were constructed. The effect of LvNELL1 infection on the proliferation, osteogenesis, and adipogenesis of human adipose-derived stem cells (hADSCs) in vitro was assessed and compared with that of LvBMP2. The results showed that hADSCs infected with LvNELL1 could efficiently and stably overexpress the target genes. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay results demonstrated that LvBMP2, but not LvNELL1, enhanced the proliferation of hADSCs. Assessment of alkaline phosphatase activity and cellular mineralization indicated that LvNELL1 infection promoted the osteogenic differentiation of hADSCs, and the effect was comparable with that of LvBMP2. Real-time polymerase chain reaction (PCR) revealed that LvNELL1 infection upregulated OSX expression but not RUNX2 expression in hADSCs. In addition, adipogenic markers (lipid droplets, peroxisome proliferator-activating receptor γ, and lipoprotein lipase) analysis showed that LvNELL1 could dramatically inhibit the adipogenic differentiation of hADSCs, but LvBMP2 had no such effect. Taken together, these findings suggested that lentiviral-mediated NELL1 gene transfer in hADSCs may be a novel and promising approach to achieve effective and precise bone regeneration.
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