Comparison of Non-Viral Transfection Methods in Melanoma Cell Primary Cultures

Micka, Bettina; Trojaneck, Beate; Niemitz, Sigrun; Lefterova, Petja; Kruopis, Susanne; Huhn, D.; Wittig, Burghardt; Schadendorf, Dirk; Schmidt‐Wolf, Ingo G.H.

doi:10.1006/cyto.1999.0621

Cited by 7 publications

(6 citation statements)

References 9 publications

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“…However, studies of other lipofection agents, including PrimeFectort (EquiBio, Ashford, UK), EasyFectort (EquiBio) and SuperFectt (Qiagen) have yielded transfection efficiencies lower than those of electroporation and LipofectAMINEt. 46 In the present study, the efficiency of PDBA-mediated gene transfer was 21.4%, which is similar to that of LipofectAMINEt. Therefore, PDBA-mediated gene transfer appears to be a safe and effective method.…”

Section: Discussionsupporting

confidence: 82%

“…Non-viral vectors may be good clinical options because they are safer and less labor intensive than virus-based methods; however, present non-viral transfection efficiencies need to be improved. Bettina et al 46 reported a 2379% transfection efficiency for LipofectAMINEt (Invitrogen)-based transfection of primary malignant lymphoma cells with vector expressing the IL-2 gene. However, studies of other lipofection agents, including PrimeFectort (EquiBio, Ashford, UK), EasyFectort (EquiBio) and SuperFectt (Qiagen) have yielded transfection efficiencies lower than those of electroporation and LipofectAMINEt.…”

Section: Discussionmentioning

confidence: 99%

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Antitumor effects of the MIG and IP-10 genes transferred with poly [D,L-2,4-diaminobutyric acid] on murine neuroblastoma

et al. 2007

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The number of tumor-infiltrating lymphocytes is known to be related to outcomes in patients with a variety of malignancies. Interferon (IFN) g-inducible protein-10 (IP-10) and monokine induced by IFNg (MIG) have chemotactic effects on activated T lymphocytes and natural killer (NK) cells. The aim of this study was to evaluate the antitumor effects of exogenous expression of the MIG and IP-10 genes delivered to solid tumors by poly [D,L-2,4-diaminobutyric acid] (PDBA). The murine MIG and IP-10 genes were transfected into mouse neuroblastoma cells with PDBA. MIG and IP-10 levels in supernatants of transfected cells were measured by enzyme-linked immunosorbent assay. The chemotactic activities of MIG and IP-10 in the supernatants of cell cultures were measured by chemotaxis assay. Tumors were injected in vivo with PDBA/pmMIGHIP-10 complexes to evaluate the effects of these genes on tumor volume and survival time of mice. Transfected PDBA/pmMIGHIP-10 complexes produced MIG and IP-10 protein in vitro. MIG and IP-10 proteins secreted into the culture medium showed chemotactic activity. MIG and IP-10 gene therapy with the PDBA system in vivo significantly inhibited tumor growth and prolonged survival time of mice. In conclusion, PDBA-mediated MIG and IP-10 gene therapy may be useful for treatment of solid tumors.

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Section: Discussionsupporting

confidence: 82%

Section: Discussionmentioning

confidence: 99%

Antitumor effects of the MIG and IP-10 genes transferred with poly [D,L-2,4-diaminobutyric acid] on murine neuroblastoma

et al. 2007

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“…[1][2][3][4][5][6][7] It is an attractive approach for gene therapy as it is the most efficient among nonviral methods. [8][9][10] In contrast with virus-based transfection, DNA delivered by electroporation is not fully optimized with respect to overcoming cellular barriers such as the plasma membrane, cytoplasmic crowding, or the nuclear envelope. However, electroporation has the advantage of being safer because complications such as viral infections cannot occur.…”

Section: Introductionmentioning

confidence: 99%

Intracellular Tracking of Single-plasmid DNA Particles After Delivery by Electroporation

et al. 2013

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Electroporation is a physical method of transferring molecules into cells and tissues. It takes advantage of the transient permeabilization of the cell membrane induced by electric field pulses, which gives hydrophilic molecules access to the cytoplasm. This method offers high transfer efficiency for small molecules that freely diffuse through electrically permeabilized membranes. Larger molecules, such as plasmid DNA, face several barriers (plasma membrane, cytoplasmic crowding, and nuclear envelope), which reduce transfection efficiency and engender a complex mechanism of transfer. Our work provides insight into the way electrotransferred DNA crosses the cytoplasm to reach the nucleus. For this purpose, single-particle tracking experiments of fluorescently labeled DNA were performed. Investigations were focused on the involvement of the cytoskeleton using drugs disrupting or stabilizing actin and tubulin filaments as the two relevant cellular networks for particle transport. The analysis of 315 movies (~4,000 trajectories) reveals that DNA is actively transported through the cytoskeleton. The large number of events allows a statistical quantification of the DNA motion kinetics inside the cell. Disruption of both filament types reduces occurrence and velocities of active transport and displacements of DNA particles. Interestingly, stabilization of both networks does not enhance DNA transport.

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“…Non-viral methods include the use of electroporation and liposomes. These methods have been compared without consistent demonstration of superior transfection efficiency [27,28]. Viral and non-viral vectors have also been compared and similar levels of transfection efficiency were seen, [29] though the clinical efficacy and immunogenicity of viral and non-viral vectors have not been directly compared.…”

Section: Whole Cell-only Strategies: Genetic Modificationmentioning

confidence: 99%