The use of an in vitro 3D melanoma model to predict in vivo plasmid transfection using electroporation

Marrero, Bernadette; Heller, Richard

doi:10.1016/j.biomaterials.2011.12.049

Cited by 45 publications

(21 citation statements)

References 38 publications

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“…Small molecules can be efficiently transferred into cells, including the ones present inside the spheroids, gene expression is limited to the external layers of cells (35). Taken together, these results, in agreement with the ones obtained by the group of R. Heller (36), indicate that the spheroid model is more relevant to an in vivo situation than cells cultured as monolayers (37,38).…”

Section: Lipid Vesicles and 3d Cell Cultures As Other Models To supporting

confidence: 87%

How Imaging Molecule Uptake into Cells can Reveal the Mechanisms of Membrane Electropermeabilization

Rols

2016

IFMBE Proceedings

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Abstract-Cell membranes can be transiently permeabilized under application of electric field pulses. This process, called electropermeabilization or electroporation, allows hydrophilic molecules, such as anticancer drugs and DNA, to enter into cells and tissues. Single cell imaging experiments revealed that the uptake of molecules is a complex multi-step that takes place in well-defined membrane regions under processes that affect the entire membrane, depend on the properties of the molecules (size, charge) and in which the cytoskeleton can play a major role.

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Section: Lipid Vesicles and 3d Cell Cultures As Other Models To supporting

confidence: 87%

How Imaging Molecule Uptake into Cells can Reveal the Mechanisms of Membrane Electropermeabilization

Rols

2016

IFMBE Proceedings

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“…It is important to develop and assess the efficacy of a transfection protocol in 3-D cell culture systems before using it to transfect cells in 3-D culture systems for invitro applications 51, 52 , or before using it for in-vivo transfection 53 .…”

Section: Discussionmentioning

confidence: 99%

Transfection in the third dimension

Dhaliwal¹,

Oshita²,

Segura³

2013

Integr. Biol.

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An understanding of parameters that modulate gene transfer in 3-D will assist in the formation of gene delivery systems and scaffolds, which can mediate efficient non-viral delivery for guiding in-vivo tissue regeneration and therapy. We have previously demonstrated the cell area and length, integrin expression, and RhoGTPases mediated signalling to be pivotal parameters that guide gene transfer to mouse mesenchymal stem cells (mMSCs) cultured in 2-D and are modulated by ECM proteins. In this study, we were interested in determining if cationic polymer mediated gene transfer to cells seeded in 3-D would occur through different mechanisms as compared to 2-D. In particular, we examined the endocytosis pathways used to internalize polyplexes, and the role of cytoskeletal dynamics and RhoGTPases on non-viral gene transfer for cells seeded in 2-D and 3-D. Inhibition of clathrin- and caveolae- mediated endocytosis resulted in more drastic decrease in overall transgene expression for cells seeded in 3-D than those in 2-D. In addition, polyplex internalization was only significantly decreased in 3-D when clathrin-mediated endocytosis was inhibited, while caveolae-mediated endocytosis inhibition for cells seeded in 2-D resulted in the strongest polyplex internalization inhibition. Actin and microtubule polymerization affected 2-D and 3-D transfection differently. Microtubule depolymerization enhanced transgene expression in 2-D, but inhibited transgene expression in 3-D. Last, inhibition of RhoGTPases also affected 2-D and 3-D transfection differently. The inhibition of ROCK effector resulted in a decrease of transgene expression and internalization for cells seeded in 3-D, but not 2-D and the inhibition of effector PAK1 resulted in an increase of transgene expression for both 2-D and 3-D. Overall, our study suggests that the process of gene transfer occurs through different mechanisms for cells seeded in 2-D compared to those seeded in 3-D.

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“…[40] It represents ap owerful tool for ab etter understanding of the influence of the cellular microenvironment on tumor biology.A s shown in Figure S16, G1-F7 4.9 and G2-F7 11.3 exhibit much higher efficacies on HEK293 3D spheroids than Lipo 2000. We further compare the efficacies of fluorodendrimers and Lipo 2000 on 3D spheroids.Unlike conventional cells cultured in am onolayer,t he 3D cell culture preserves specific biochemical and morphological features of corresponding tissues in vivo.T he use of 3D cell culture can predict in vivo gene transfection efficacy of am aterial.…”

Section: Methodsmentioning

confidence: 99%

Self‐Assembled Fluorodendrimers Combine the Features of Lipid and Polymeric Vectors in Gene Delivery

Wang

et al. 2015

Angewandte Chemie

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An ideal vector in gene therapyshould exhibit high serum stability,e xcellent biocompatibility,adesired transfection efficacy and permeability into targeted tissues.Here,we describe aclass of low-molecular-weight fluorodendrimers for efficient gene delivery.T hese materials self-assemble into uniform nanospheres and allow for efficient transfection at low charge ratios and very low DNAd oses with minimal cytotoxicity.O ur results demonstrate that these vectors combine the features of synthetic gene vectors such as liposomes and cationic polymers and present promising potential for clinical gene therapy.

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The use of an in vitro 3D melanoma model to predict in vivo plasmid transfection using electroporation

Cited by 45 publications

References 38 publications

How Imaging Molecule Uptake into Cells can Reveal the Mechanisms of Membrane Electropermeabilization

How Imaging Molecule Uptake into Cells can Reveal the Mechanisms of Membrane Electropermeabilization

Transfection in the third dimension

Self‐Assembled Fluorodendrimers Combine the Features of Lipid and Polymeric Vectors in Gene Delivery

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