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Weiwei Wang,
Wenzhong Li,
Lailiang Ou,
Eva Flick,
Peter Mark, Catharina Nesselmann,
Cornelia A Lux,
Hans-Heinrich Gatzen,
Alexander Kaminski,
Andreas Liebold,
Karola Lützow,
Andreas Lendlein,
Ren-Ke Li,
Gustav Steinhoff,
Nan Ma
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ABSTRACT: Transplantation of mesenchymal stem cells (MSCs) derived from adult bone marrow has been proposed as a potential therapeutic approach for post-infarction left ventricular (LV) dysfunction. However, age-related functional decline of stem cells has restricted their clinical benefits after transplantation into the infarcted myocardium. The limitations imposed on patient cells could be addressed by genetic modification of stem cells. This study was designed to improve our understanding of genetic modification of human bone marrow derived mesenchymal stem cells (hMSCs) by polyethylenimine (PEI, branched with Mw 25 kD), one of non-viral vectors that show promise in stem cell genetic modification, in the context of cardiac regeneration for patients. We optimized the PEI-mediated reporter gene transfection into hMSCs, evaluated whether transfection efficiency is associated with gender or age of the cell donors, analysed the influence of cell cycle on transfection and investigated the transfer of therapeutic vascular endothelial growth factor gene (VEGF). hMSCs were isolated from patients with cardiovascular disease aged from 41 to 85 years. Optimization of gene delivery to hMSCs was carried out based on the particle size of the PEI/DNA complexes, N/P ratio of complexes, DNA dosage and cell viability. The highest efficiency with the cell viability near 60% was achieved at N/P ratio 2 and 6.0 μg DNA/cm(2) . The average transfection efficiency for all tested samples, middle-age group (<65 years), old-age group (>65 years), female group and male group was 4.32%, 3.85%, 4.52%, 4.14% and 4.38%, respectively. The transfection efficiency did not show any correlation either with the age or the gender of the donors. Statistically, there were two subpopulations in the donors; and transfection efficiency in each subpopulation was linearly related to the cell percentage in S phase. No significant phenotypic differences were observed between these two subpopulations. Furthermore, PEI-mediated therapeutic gene VEGF transfer could significantly enhance the expression level.
Journal of Cellular and Molecular Medicine 09/2011; 15(9):1989-98. · 4.13 Impact Factor
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ABSTRACT: Accumulating clinical and experimental evidence indicates that stem cells from various sources are promising in the treatment of cardiac dysfunction. They may be incorporated into neovascular foci and thus contribute to postnatal physiological and pathological vasculogenesis and/or produce a variety of growth factors for angiogenesis and cytokines that home other stem cells from other organs for cardiac regeneration. This review focuses on the neovascularization of stem cells from different sources in cardiac repair, with emphasis on adult stem cells.
Therapeutic Advances in Cardiovascular Disease 02/2010; 4(1):27-42.
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ABSTRACT: Accumulating clinical and experimental evidence indicates that mesenchymal stem cells (MSCs) are promising cell types in the treatment of cardiac dysfunction. They may trigger production of reparative growth factors, replace damaged cells and create an environment that favours endogenous cardiac repair. However, identifying mechanisms which regulate the role of MSCs in cardiac repair is still at work. To achieve the maximal clinical benefits, ex vivo manipulation can further enhance MSC therapeutic potential. This review focuses on the mechanism of MSCs in cardiac repair, with emphasis on ex vivo manipulation.
Journal of Cellular and Molecular Medicine 09/2008; 12(5B):1795-810. · 4.13 Impact Factor
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Wenzhong Li,
Ph.D. Nan Ma M.D,
Lee-Lee Ong, Catharina Nesselmann,
Christian Klopsch,
Yury Ladilov,
Dario Furlani,
Christoph Piechaczek,
Jeannette M. Moebius,
Karola Lützow,
Andreas Lendlein,
Christof Stamm,
Ren-Ke Li,
Gustav Steinhoff
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ABSTRACT: Engraftment of mesenchymal stem cells (MSCs) derived from adult bone marrow has been proposed as a potential therapeutic approach for postinfarction left ventricular dysfunction. However, limited cell viability after transplantation into the myocardium has restricted its regenerative capacity. In this study, we genetically modified MSCs with an antiapoptotic Bcl-2 gene and evaluated cell survival, engraftment, revascularization, and functional improvement in a rat left anterior descending ligation model via intracardiac injection. Rat MSCs were manipulated to overexpress the Bcl-2 gene. In vitro, the antiapoptotic and paracrine effects were assessed under hypoxic conditions. In vivo, the Bcl-2 gene-modified MSCs (Bcl-2-MSCs) were injected after myocardial infarction. The surviving cells were tracked after transplantation. Capillary density was quantified after 3 weeks. The left ventricular function was evaluated by pressure-volume loops. The Bcl-2 gene protected MSCs against apoptosis. In vitro, Bcl-2 overexpression reduced MSC apoptosis by 32% and enhanced vascular endothelial growth factor secretion by more than 60% under hypoxic conditions. Transplantation with Bcl-2-MSCs increased 2.2-fold, 1.9-fold, and 1.2-fold of the cellular survival at 4 days, 3 weeks, and 6 weeks, respectively, compared with the vector-MSC group. Capillary density in the infarct border zone was 15% higher in Bcl-2-MSC transplanted animals than in vector-MSC treated animals. Furthermore, Bcl-2-MSC transplanted animals had 17% smaller infarct size than vector-MSC treated animals and exhibited functional recovery remarkably. Our current findings support the premise that transplantation of antiapoptotic gene-modified MSCs may have values for mediating substantial functional recovery after acute myocardial infarction.
Stem Cells 07/2007; 25(8):2118 - 2127. · 7.78 Impact Factor
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ABSTRACT: Molecular Therapy (2006) 13, S169|[ndash]|S169; doi: 10.1016/j.ymthe.2006.08.507
440. Overexpression of Bcl-2 Prevented MSCs from Apoptosis and Improved Heart Function
Nan Ma1, Wenzhong Li1, Catharina Nesselmann1, Leelee Ong1, Alexander Kaminski1, Brigitte M. P|[uuml]|tzer2, Ren-Ke Li3, Christof Stamm1 and Gustav Steinhoff11Department of Cardiac Surgery, University Rostock, Rostock, Germany2Department of Vectorology and Experimental Gene Therapy, University Rostock, Rostock, Germany3Division of Cardiovascular Surgery, Toronto General Hospital and the University of Toronto, Toronto, Canada
Molecular Therapy 04/2006; · 6.87 Impact Factor
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Wenzhong Li, Catharina Nesselmann,
Zhaohui Zhou,
Lee-Lee Ong,
Ferenc Öri,
Guping Tang,
Alexander Kaminski,
Karola Lützow,
Andreas Lendlein,
Andreas Liebold,
Christof Stamm,
John Wang,
Gustav Steinhoff,
Nan Ma
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ABSTRACT: Gene delivery with non-viral gene vectors to the cardiovascular system suffers from low transfection efficiency. In this study, magnetic fields were investigated to assist cardiovascular gene delivery via magnetic nanobeads both in vitro and in vivo. The magnetic field was provided with a 1120 mT Nd–Fe–B permanent magnet while complexes of poly-ethyleneimine (PEI) and various DNA plasmids were conjugated with magnetic nanobeads (MNB) using a Sulfo-NHS-LC-Biotin linker. In vitro results showed that transfection in two cell lines was 30–80-fold higher in magnetically conjugated MNB/PEI/DNA complexes than transfection from PEI/DNA complexes alone. Similarly, in vivo results using mouse models showed 72 h after injection observable gene expression in the heart with conjugated MNB/PEI/DNA complexes, but barely with PEI/DNA complexes alone.
Journal of Magnetism and Magnetic Materials 311(1):336-341. · 1.78 Impact Factor
