Polyethylenimine-mediated gene delivery into human bone marrow mesenchymal stem cells from patients
| Autor: | Wang, Weiwei, Li, Wenzhong, Ou, Lailiang, Flick, Eva, Mark, Peter, Nesselmann, Catharina, Lux, Cornelia A., Gatzen, Hanz-Heinrich, Kaminski, Alexander, Liebold, Andreas, Lützow, Karola, Lendlein, Andreas, Li, Ren-Ke, Steinhoff, Gustav, Ma, Nan |
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| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
green fluorescent protein
Adult Male Vascular Endothelial Growth Factor A phenotype Cell Survival Green Fluorescent Proteins Bone Marrow Cells Polyethylenimine Transfection S Phase VEGFA protein human middle aged Humans Polyethyleneimine ddc:610 Gene delivery human gene transfer mesenchymal stem cell Aged Aged 80 and over Cell Death drug effect article Gene Transfer Techniques Mesenchymal Stem Cells DNA Cardiovascular disease Dewey Decimal Classification::600 | Technik cell cycle S phase female Non-viral vector vasculotropin A Human bone marrow derived mesenchymal stem cells cytology genetic transfection Dewey Decimal Classification::600 | Technik::610 | Medizin Gesundheit ddc:600 metabolism bone marrow cell |
| Zdroj: | Journal of Cellular and Molecular Medicine 15 (2011), Nr. 9 |
| Popis: | 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), 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. DFG/SFB/Transregio 37 BMBF/0313191 German Helmholtz Association DFG/0402710 Ministry of Education/0312138 A Ministry of Economy (Mecklenburg-West Pommerania)/V220-630-08-TFMVF/S-035 Marie Curie International Research Staff Exchange Scheme (IRSES, FP7-PEOPLE-2009-IRSES) Reference and Translation Center for Cardiac Stem Cell Therapy (RTC) |
| Databáze: | OpenAIRE |
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