Polyethylenimine-mediated gene delivery into human bone marrow mesenchymal stem cells from patients
Creators: name: Wang, Weiwei
name: Li, Wenzhong
name: Ou, Lailiang
name: Flick, Eva
name: Mark, Peter
name: Nesselmann, Catharina
name: Lux, Cornelia A.
name: Gatzen, Hanz-Heinrich
name: Kaminski, Alexander
name: Liebold, Andreas
name: Lützow, Karola
name: Lendlein, Andreas
name: Li, Ren-Ke
name: Steinhoff, Gustav
name: Ma, Nan
Publisher: name: Chichester : Wiley-Blackwell Publishing Ltd.
Year: 2011
Subjects: subject: Cardiovascular disease
subject: Gene delivery
subject: Human bone marrow derived mesenchymal stem cells
subject: Non-viral vector
subject: Polyethylenimine
subject: DNA
subject: green fluorescent protein
subject: polyethyleneimine
subject: vasculotropin A
subject: VEGFA protein, human
subject: adult
subject: aged
subject: article
subject: bone marrow cell
subject: cell cycle S phase
subject: cell death
subject: cell survival
subject: cytology
subject: drug effect
subject: female
subject: gene transfer
subject: genetic transfection
subject: human
subject: male
subject: mesenchymal stem cell
subject: metabolism
subject: middle aged
subject: phenotype
subject: Adult
subject: Aged
subject: Aged, 80 and over
subject: Bone Marrow Cells
subject: Cell Death
subject: Cell Survival
subject: Female
subject: Gene Transfer Techniques
subject: Green Fluorescent Proteins
subject: Humans
subject: Male
subject: Mesenchymal Stem Cells
subject: Middle Aged
subject: Phenotype
subject: Polyethyleneimine
subject: S Phase
subject: Transfection
subject: Vascular Endothelial Growth Factor A
subject: Dewey Decimal Classification::600 | Technik
subject: Dewey Decimal Classification::600 | Technik::610 | Medizin, Gesundheit

description: 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 (&lt;65 years), old-age group (&gt;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.