Cell line-dependent internalization pathways determine DNA transfection efficiency of decaarginine-PEG-lipid

Izumisawa, Tomohiro; Hattori, Yoshiyuki; Date, Masataka; Toma, Kazunori; Maitani, Yoshie

doi:10.1016/j.ijpharm.2010.11.017

Cited by 24 publications

(14 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Vector complex size is known to affect the mechanism of cell internalization [51,52] and plays an important role in endocytosis and migration of the expression package to the nucleus [23], with cytoplasmic mobility and nuclear entry inversely proportional to the size of the plasmid DNA [53]. Since minicircle plasmids form smaller complexes with cationic lipids, such as Lipofectamine, these smaller minicircle complexes may interact more efficiently with the cell membrane, leading to an improved cellular uptake [48], and enhanced intracellular trafficking and nuclear entry [24].…”

Section: Resultsmentioning

confidence: 99%

“…In mammalian cells the mechanisms of cellular attachment and internalisation of DNA-lipid complexes following transfection, has been shown to be cell line-dependant [51,55]. Studies investigating transfection efficiencies of human cervical carcinoma (HeLa) cells, African green monkey kidney fibroblast-like cell line (COS-7), Chinese hamster ovary (CHO) and HEK293T cell lines have shown differences in their internalisation pathways, which have resulted in variable transfection efficiency [51,55].…”

Section: Resultsmentioning

confidence: 99%

“…Studies investigating transfection efficiencies of human cervical carcinoma (HeLa) cells, African green monkey kidney fibroblast-like cell line (COS-7), Chinese hamster ovary (CHO) and HEK293T cell lines have shown differences in their internalisation pathways, which have resulted in variable transfection efficiency [51,55]. It is possible that equine MSCs exhibit a similar effect on the subcelluar machinery required for effective internalisation, compared to canine and rat MSCs, affecting cellular uptake and intracellular trafficking of plasmid motifs and therefore transfection efficiency.…”

Section: Resultsmentioning

confidence: 99%

See 2 more Smart Citations

Minicircle Mediated Gene Delivery to Canine and Equine Mesenchymal Stem Cells

Tidd

Michelsen

Hilbert

et al. 2017

IJMS

View full text Add to dashboard Cite

Gene-directed tissue repair offers the clinician, human or veterinary, the chance to enhance cartilage regeneration and repair at a molecular level. Non-viral plasmid vectors have key biosafety advantages over viral vector systems for regenerative therapies due to their episomal integration however, conventional non-viral vectors can suffer from low transfection efficiency. Our objective was to identify and validate in vitro a novel non-viral gene expression vector that could be utilized for ex vivo and in vivo delivery to stromal-derived mesenchymal stem cells (MSCs). Minicircle plasmid DNA vector containing green fluorescent protein (GFP) was generated and transfected into adipose-derived MSCs from three species: canine, equine and rodent and transfection efficiency was determined. Both canine and rat cells showed transfection efficiencies of approximately 40% using minicircle vectors with equine cells exhibiting lower transfection efficiency. A Sox9-expressing minicircle vector was generated and transfected into canine MSCs. Successful transfection of the minicircle-Sox9 vector was confirmed in canine cells by Sox9 immunostaining. This study demonstrate the application and efficacy of a novel non-viral expression vector in canine and equine MSCs. Minicircle vectors have potential use in gene-directed regenerative therapies in non-rodent animal models for treatment of cartilage injury and repair.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Minicircle Mediated Gene Delivery to Canine and Equine Mesenchymal Stem Cells

Tidd

Michelsen

Hilbert

et al. 2017

IJMS

View full text Add to dashboard Cite

show abstract

“…In epithelial cells, filomicelles (PEGylated by definition) undergo fragmentation upon macropinocytosis [127]. It should be noted, not surprisingly, that the very same PEGylation that enhances circulation (see ‘Circulation’ section) can impact internalization and subcellular trafficking to varying extents (sometimes negatively) [133,134]. Therefore, while several studies have discussed the complex interplay of surface chemistry (e.g., PEGylation) and nanocarrier surface binding moiety affinity (e.g., antibodies), nanocarrier geometry is an emerging approach that also modulates uptake and intracellular addressing of carrier cargo, which may be used for prolongation of effects of antioxidant enzymes.…”

Section: Modulation Of Geometry Of Polymeric Carriers For Antioxidantsmentioning

confidence: 99%

Nanocarriers for Vascular Delivery of Antioxidants

et al. 2011

View full text Add to dashboard Cite

Antioxidant enzymes (AOEs) catalase and superoxide dismutase (SOD) detoxify harmful reactive oxygen species, but the therapeutic utility of AOEs is hindered by inadequate delivery. AOE modification by polyethylene glycol (PEG) and encapsulation in PEG-coated liposomes increases the AOE bioavailability and enhances protective effects in animal models. Pluronic-based micelles formed with AOEs show even more potent protective effects. Furthermore, polymeric nanocarriers (PNCs) based on PEG-copolymers protect encapsulated AOEs from proteolysis and improve delivery to the target cells, such as the endothelium lining the vascular lumen. Antibodies to endothelial determinants conjugated to AOEs or AOE carriers provide targeting and intracellular delivery. Targeted liposomes, protein conjugates and magnetic nanoparticles deliver AOEs to sites of vascular oxidative stress in the cardiovascular, pulmonary and nervous systems. Further advances in nanodevices for AOE delivery will provide a basis for the translation of this approach in the clinical domain.

show abstract

“…They demonstrated different TE and even the same cell line type, but from different subjects, also had different TE. Cell physiology might be a critical factor in cellular internalization and transfection [36]. Further investigation is required to assist in providing an explanation.…”

Section: Discussionmentioning

confidence: 99%

Ultrasound Treatment Increases Transfection Efficiency of Low Molecular Weight Chitosan in Fibroblasts but Not in KB Cells

2014

View full text Add to dashboard Cite

The aim of this study was to optimize transfection efficiency (TE) of the depolymerized low molecular weight (LW) chitosan with molecular weight (Mw) at 16 kDa and 54% degree of deacetylation (DDA) on three primary cells of fibroblast (F), dental pulp (P), and periodontal ligament (PDL). The effect of low frequency ultrasound treatment on the chitosan-DNA complexes prior transfection on TE was also evaluated. This LW chitosan required high N/P ratio (>34) to bind DNA completely. An N/P ratio above 56 tended to improve TE in most primary cells nearly at the level of Lipofectamine. Ultrasonication can reduce the aggregation and sizes of the chitosan-DNA microparticles. It increased TE of F cells at an N/P ratio above 34, which was higher than Lipofectamine. However, this ultrasound treatment caused loss of TE in KB cells. MTT assay of these chitosan-DNA complexes revealed no significant cytotoxicity to both KB and F cells. This LW chitosan has potential for further development into a safer alternative to gene delivery systems in various cells of interest; however the optimal conditions have to be adjusted, depending on each cell source.

show abstract

Cell line-dependent internalization pathways determine DNA transfection efficiency of decaarginine-PEG-lipid

Cited by 24 publications

References 45 publications

Minicircle Mediated Gene Delivery to Canine and Equine Mesenchymal Stem Cells

Minicircle Mediated Gene Delivery to Canine and Equine Mesenchymal Stem Cells

Nanocarriers for Vascular Delivery of Antioxidants

Ultrasound Treatment Increases Transfection Efficiency of Low Molecular Weight Chitosan in Fibroblasts but Not in KB Cells

Contact Info

Product

Resources

About