Polyethylene Glycol-Conjugated Copolymers for Plasmid DNA Delivery

Lee, Minhyung; Kim, Sung Wan

doi:10.1007/s11095-004-9003-5

Cited by 253 publications

(152 citation statements)

References 60 publications

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“…Such difference might be the result of difference in GFP expression in hUSSCs and hMSCs or size-dependency of nucleic acid transfection in both stem cell types. Altogether it revealed that the nature of the cell itself plays a critical role in effectiveness of transfection as suggested by others (Yamano et al 2010;Lee and Kim 2005).…”

Section: Discussionsupporting

confidence: 54%

A comparative study on nonviral genetic modifications in cord blood and bone marrow mesenchymal stem cells

et al. 2012

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The focus of both clinical and basic studies on stem cells is increasing due to their potentials in regenerative medicine and cell-based therapies. Recently stem cells have been genetically modified to enhance an existing character in or to bring a new property to them. However, accomplishment of declared goals requires detailed knowledge about their molecular characteristics which could be achieved by genetic modifications mostly through nonviral transfection strategies. Capable of differentiating into multiple cells, human unrestricted somatic stem cells (hUSSCs) and human mesenchymal stem cells (hMSCs) seem to be suitable candidates for transfection approaches. Involvement of microRNAs (miRNAs) in many biological processes makes their transfection evaluation valuable. Herein we investigated the efficacy and toxicity of four typically used transfection reagents (Arrest-In, Lipofectamine 2000, Oligofectamine and HiPerfect) systematically to deliver fluorescent labeled-miRNA and Green Fluorescent Protein (GFP) expressing plasmid into hUSSCs and hMSCs. The authenticity of stem cells was verified by differentiation experiments along with flow cytometry of surface markers. Our study revealed that stemness properties of these stem cells were not affected by transient transfection. Moreover the ratios of cell viability and transfection efficiency in both analyzed stem cells were reversed. Considering cell viability, the highest fraction of GFP-expressing cells was obtained using Oligofectamine (*50%) while the highest transfection rate of miRNA was achieved by Lipofectamine 2000 (*90%). Moreover dependency of hMSCs to size of transfected nucleic acid and timedependency of Oligofectamine and their affection on the yield of transfection were observed. Cytotoxicity assessments also showed that hUSSCs are sensitive to HiPerFect. In addition cells treated by Lipofectamine showed morphological changes. Representing the efficient nucleic acid transfection, our research facilitates comprehensive genetic modification of stem cells and demonstrates powerful approaches to understand stem cell molecular regulation mechanisms, Electronic supplementary material The online version of this article

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Section: Discussionsupporting

confidence: 54%

A comparative study on nonviral genetic modifications in cord blood and bone marrow mesenchymal stem cells

et al. 2012

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“…Finally, PEG-graftment has been successfully used as a spacer moiety between a targeting ligand and a cationic polymer. 21 Moreover, a phase I clinical trial using PEGylated PLL/pDNA complexes has been conducted in cystic fibrosis (CF) patients with some success in partial correction of the CF chloride transport defect, while no inflammatory response or other toxicity due to the gene vector was observed. 33 Nevertheless, an ideal nonviral gene transfer agent has not been identified up to now.…”

Section: Introductionmentioning

confidence: 99%

Characterization of Tailor-Made Copolymers of Oligo(ethylene glycol) Methyl Ether Methacrylate and N,N-Dimethylaminoethyl Methacrylate as Nonviral Gene Transfer Agents: Influence of Macromolecular Structure on Gene Vector Particle Properties and Transfection Efficiency

et al. 2009

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Oligo(ethylene glycol) methyl ether methacrylates (OEGMA) of various chain lengths (i.e., 9, 23, or 45 EG units) and N,N-dimethylaminoethyl methacrylate (DMAEMA) were copolymerized by atom transfer radical polymerization (ATRP), yielding well-defined P(DMAEMA-co-OEGMA) copolymers with increasing OEGMA molar fractions (F OEGMA) but a comparable degree of polymerization (DP ∼ 120). Increase of both F OEGMA and OEGMA chain lengths correlated inversely with gene vector size, morphology, and zeta potential. P(DMAEMA-co-OEGMA) copolymers prevented gene vector aggregation at high plasmid DNA (pDNA) concentrations in isotonic solution and did not induce cytotoxicity even at high concentrations. Transfection efficiency of the most efficient P(DMAEMA-co-OEGMA) copolymers was found to be >10-fold lower compared with branched polyethylenimine (PEI) 25 kDa. Although OEGMA copolymerization largely reduced gene vector binding with the cell surface, cellular internalization of the bound complexes was less affected. These observations suggest that inefficient endolysosomal escape limits transfection efficiency of P(DMAEMA-co-OEGMA) copolymer gene vectors. Despite this observation, optimized p(DMAEMA-co-OEGMA) gene vectors remained stable under conditions for in vivo application leading to 7-fold greater gene expression in the lungs compared with PEI. Tailor-made P(DMAEMA-co-OEGMA) copolymers are promising nonviral gene transfer agents that fulfill the requirements for successful in vivo gene delivery.

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“…Furthermore, polyethylene glycosylation (PEGylation) can prevent salt-induced aggregation through steric stabilization [25,26,[29][30][31]33,34]. Additionally, PEG is often used as a spacer for targeting ligands since the shielding effect of PEG is able to decrease nonspecific interactions with negatively charged cellular membranes, which results in reduction of nonspecific cellular uptake [37]. While some PEGylation strategies have had no effect on transfection efficiency in vitro [25,31,33] or in vivo [25], or even enhanced transfection [27,34], others have reported that PEGylation resulted in poor transfection [28,29,32], presumably due to interference with complexation [36].…”

Section: Introductionmentioning

confidence: 99%

Surface polyethylene glycol enhances substrate-mediated gene delivery by nonspecifically immobilized complexes

2008

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Substrate-mediated gene delivery describes the immobilization of gene therapy vectors to a biomaterial, which enhances gene transfer by exposing adhered cells to elevated DNA concentrations within the local microenvironment. Surface chemistry has been shown to affect transfection by nonspecifically immobilized complexes using self-assembled monolayers (SAMs) of alkanethiols on gold. In this report, SAMs were again used to provide a controlled surface to investigate whether the presence of oligo(ethylene glycol) (EG) groups in a SAM could affect complex morphology and enhance transfection. EG groups were included at percentages that did not affect cell adhesion. Nonspecific complex immobilization to SAMs containing combinations of EG-and carboxylic acidterminated alkanethiols resulted in substantially greater transfection than surfaces containing no EG groups or SAMs composed of EG groups combined with other functional groups. Enhancement in transfection levels could not be attributed to complex binding densities or release profiles. Atomic force microscopy imaging of immobilized complexes revealed that EG groups within SAMs affected complex size and appearance and could indicate the ability of these surfaces to preserve complex morphology upon binding. The ability to control the morphology of the immobilized complexes and influence transfection levels through surface chemistry could be translated to scaffolds for gene delivery in tissue engineering and diagnostic applications.

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Polyethylene Glycol-Conjugated Copolymers for Plasmid DNA Delivery

Cited by 253 publications

References 60 publications

A comparative study on nonviral genetic modifications in cord blood and bone marrow mesenchymal stem cells

A comparative study on nonviral genetic modifications in cord blood and bone marrow mesenchymal stem cells

Characterization of Tailor-Made Copolymers of Oligo(ethylene glycol) Methyl Ether Methacrylate and N,N-Dimethylaminoethyl Methacrylate as Nonviral Gene Transfer Agents: Influence of Macromolecular Structure on Gene Vector Particle Properties and Transfection Efficiency

Surface polyethylene glycol enhances substrate-mediated gene delivery by nonspecifically immobilized complexes

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