Determination of osteoprogenitor-specific promoter activity in mouse mesenchymal stem cells by recombinant adeno-associated virus transduction

Sustained and promoter dependent bone morphogenetic protein expression by rat mesenchymal stem cells after BMP-2 transgene electrotransfer

“…In contrast, the levels of hBMP-2 expression driven by the osteocalcin promoter markedly increased over the same time period and correlated to the observed increased BMP-2 production rates; in fact, the production rates driven by the osteocalcin promoter were statistically equivalent to those driven by the strongest promoters (specifically, the EF-1a, GAPDH and β-actin promoters) at day 14 after transgene electrotransfer into MSCs. Transgene expression driven by the promoter of osteocalcin (a protein specifically expressed by osteoblasts) was evident only from days 7 to 21 after induction of transgenic MSC differentiation into the osteoblastic lineage (Kumar et al, 2005). The data of the present study suggest that, whereas the activity www.ecmjournal.org E Ferreira et al BMP-2 delivery by MSCs after gene electrotransfer of the osteocalcin promoter was predictably weak in undifferentiated MSCs, it might have been boosted during osteoblastic MSC differentiation, resulting in the observed up-regulation in BMP-2 expression.…”

“…The efficiency of cellular promoters has been shown to be cell specific and dependent on cell activity (Doll et al, 1996;Durieux et al, 2005;Ramezani et al, 2000;Spenger et al, 2004). While studies indicated that osteoblastic specific promoters, including the promoter of the osteocalcin gene, led to increase in transgene expression over time for BMP-2 expression in MSCs (Kumar et al, 2005), the effect of various cellular and tissue-specific promoters on the level and duration of BMP-2 expression in MSCs has not yet been well investigated. For these reasons, the present study determined the influence of the promoter driving the hBMP-2 transgene on level and duration of gene expression after transgene electrotransfer into rat MSCs.…”

Adeno-associated viral vector transduction of human mesenchymal stem cells

“…The few published studies that utilize AAV transduction of MSCs agree on the feasibility of this (Ho et al, 2004;Kumar et al, 2004;Kumar et al, 2005;Bosch et al, 2006;McMahon et al, 2006). However, these studies leave several basic questions regarding transduction efficiency and kinetics unanswered.…”

Bioluminescence imaging in bone tissue engineering

“…Using the constitutively active cytomegalovirus (CMV) promoter, cell survival and proliferation were examined both in vitro and in vivo [81,82]. The activity of several osteoprogenitor-specific promoters was assessed to monitor osteogenic differentiation of stem cells in vitro including the promoters of Runx2/Cbfa-1, osteopontin (OPN), osteocalcin (OCN), collagen type 1a (COL), alkaline phosphatise (ALP) and the bone morphogenetic protein responsive element (BRE) [83,84]. Vascularisation was evaluated by vascular endothelial growth factor receptor 2 (VEGFR2)-luciferase Chapter 1 cells and mice [55,85].…”

“…For instance, the constitutively active cytomegalovirus (CMV) promoter can be used to monitor cell survival and proliferation [81,82]. In addition, osteoblast-specific promoters have been evaluated as a tool for monitoring osteogenic differentiation including sequences derived from the Runx2/Cbfa-1 (RUNX), osteopontin (OPN), osteocalcin (OCN), collagen type 1a (COL), alkaline phosphatase (ALP) and bone morphogenetic genes [83,84]. In addition, we investigated the cellular hypoxia/nutrient status during BTE using hypoxia responsive element (HRE)-luciferase transgenic cells (Chapter 3).…”