Polyethyleneimine-mediated gene delivery into human adipose derived stem cells

Ahn, Hyun Hee; Lee, Jung Hwa; Kim, Kyung Sook; Lee, Ju Young; Kim, Moon Suk; Khang, Gilson; Lee, Il Woo; Lee, Hai Bang

doi:10.1016/j.biomaterials.2008.02.006

Cited by 83 publications

(68 citation statements)

References 32 publications

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“…MSCs are the subject of countless cell-based repair strategies either as a standalone treatment or in conjunction with scaffolds for bone regeneration [25]. PEI has been successfully used in a variety of adherent cell lines such as HeLa, HepG2 and NIH3T3 [26,27], and a marrow derived stem cell line [28]. Furthermore, Saraf et al have reported transfection efficiencies of just 12 % using branched PEI to transfect human MSCs [29].…”

Section: Introductionmentioning

confidence: 99%

“…Furthermore, Saraf et al have reported transfection efficiencies of just 12 % using branched PEI to transfect human MSCs [29]. Similarly, PEI has also been used to transfect adipose derived stem cells [26] and has even been the basis of an in vivo gene activated matrix (GAM) for bone repair [20]. It is for this reason that it was sought to determine whether PEI could provide the same transfection efficiency for pDNA in rMSCs as the targeted in vivo cell type and to also establish if PEI polyplexes could be successfully merged with our scaffolds which are specifically engineered for bone regeneration.…”

Section: Introductionmentioning

confidence: 99%

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The development of non-viral gene-activated matrices for bone regeneration using polyethyleneimine (PEI) and collagen-based scaffolds

Tierney

Duffy

Hibbitts

et al. 2012

Journal of Controlled Release

124

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The healing potential of scaffolds for tissue engineering can be enhanced by combining them with genes to produce gene-activated matrices (GAMs) for tissue regeneration. We examined the potential of using polyethyleneimine (PEI) as a vector for transfection of mesenchymal stem cells (MSCs) in monolayer culture and in 3D collagen-based GAMs. PEI-pDNA polyplexes were fabricated at a range of N/P ratios and their optimal transfection parameters (N/P 7 ratio, 2µg dose) and transfection efficiencies (45 ± 3%) determined in monolayer culture. The polyplexes were then loaded onto collagen, collagen-glycosaminoglycan and collagen-nanohydroxyapatite scaffolds where gene expression was observed up to 21 days with a polyplex dose as low as 2µg. Transient expression profiles indicated that the GAMs act as a polyplex depot system whereby infiltrating cells become transfected over time as they migrate throughout the scaffold. The collagen-nHa GAM exhibited the most prolonged and elevated levels of transgene expression. This research has thus demonstrated that PEI is a highly efficient pDNA transfection agent for both MSC monolayer cultures and in the 3D GAM environment. By combining therapeutic gene therapy with highly engineered scaffolds, it is proposed that these GAMs might have immense capability to promote tissue regeneration.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The development of non-viral gene-activated matrices for bone regeneration using polyethyleneimine (PEI) and collagen-based scaffolds

Tierney

Duffy

Hibbitts

et al. 2012

Journal of Controlled Release

124

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“…Besides the molecular weight range, N/P ratio (N = molar number of primary amines in the polymer; P = molar number of phosphate groups in the pDNA backbone) also influences transfection efficiencies. A lower N/P ratio induces higher transfection rates, but the cytotoxicity is also increased 145 . In order to increase the transfection efficiency and decrease the cytotoxicity, several studies focus on combining PEI with other biologicals with varying success.…”

Section: Synthetic Polymer-based Transfectionsmentioning

confidence: 99%

Non-viral gene therapy for bone tissue engineering

Wegman

2013

Biotechnology and Genetic Engineering Reviews

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“…[33][34][35][36] The feasibility of using polyethylenimine for transfection of many cell lines, in addition to liposomal and other lipid-based carriers, has been confirmed, 33 but several researchers have demonstrated that it is difficult for polyethylenimine to obtain higher transfection efficiency than a viral vehicle. [34][35][36] Farrell et al 34 reported that the 48-hour transfection efficiency of polyethylenimine-mediated gene delivery to rat mesenchymal stem cells is below 4% at an N/P ratio of 7. Ahn et al 35 reported that introducing DNA/PEI nanoparticles into rat mesenchymal stem cells resulted in 2%-10% transfection efficiency.…”

mentioning

confidence: 99%

“…In human adipose-derived stem cells, polyethylenimine produced the highest transfection of 19% at an N/P ratio of 8, which was superior to that achieved by Lipofectamine. 36 Cell internalization is correlated with the buffer action of the cell membrane reservoir, the functioning of the cytoskeleton, and the location of the nanomicrocapsules within cells. The interaction between nanomicrocapsules and various cell lines may be very different.…”

mentioning

confidence: 99%

Plasmid-encapsulated polyethylene glycol-grafted polyethylenimine nanoparticles for gene delivery into rat mesenchymal stem cells

Chen

Zhang²,

He³

et al. 2011

IJN

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Background: Mesenchymal stem cell transplantation is a promising method in regenerative medicine. Gene-modified mesenchymal stem cells possess superior characteristics of specific tissue differentiation, resistance to apoptosis, and directional migration. Viral vectors have the disadvantages of potential immunogenicity, carcinogenicity, and complicated synthetic procedures. Polyethylene glycol-grafted polyethylenimine (PEG-PEI) holds promise in gene delivery because of easy preparation and potentially targeting modification. Methods: A PEG8k-PEI25k graft copolymer was synthesized. Agarose gel retardation assay and dynamic light scattering were used to determine the properties of the nanoparticles. MTT reduction, wound and healing, and differentiation assays were used to test the cytobiological characteristics of rat mesenchymal stem cells, fluorescence microscopy and flow cytometry were used to determine transfection efficiency, and atomic force microscopy was used to evaluate the interaction between PEG-PEI/plasmid nanoparticles and mesenchymal stem cells. Results: After incubation with the copolymer, the bionomics of mesenchymal stem cells showed no significant change. The mesenchymal stem cells still maintained high viability, resettled the wound area, and differentiated into adipocytes and osteoblasts. The PEG-PEI completely packed plasmid and condensed plasmid into stable nanoparticles of 100–150 nm diameter. After optimizing the N/P ratio, the PEG-PEI/plasmid microcapsules delivered plasmid into mesenchymal stem cells and obtained an optimum transfection efficiency of 15%–21%, which was higher than for cationic liposomes. Conclusion: These data indicate that PEG-PEI is a valid gene delivery agent and has better transfection efficiency than cationic liposomes in mesenchymal stem cells.

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Polyethyleneimine-mediated gene delivery into human adipose derived stem cells

Cited by 83 publications

References 32 publications

The development of non-viral gene-activated matrices for bone regeneration using polyethyleneimine (PEI) and collagen-based scaffolds

The development of non-viral gene-activated matrices for bone regeneration using polyethyleneimine (PEI) and collagen-based scaffolds

Non-viral gene therapy for bone tissue engineering

Plasmid-encapsulated polyethylene glycol-grafted polyethylenimine nanoparticles for gene delivery into rat mesenchymal stem cells

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