• Title/Summary/Keyword: Magnetofection

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Magnetofection is an efficient tool for ectopic gene expression into oral cells

  • Ji, Jae-Hoon;Ko, Seon-Yle;Jang, Young-Joo
    • International Journal of Oral Biology
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    • v.32 no.1
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    • pp.7-11
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    • 2007
  • It is difficult to introduce DNA in non-invasive manner into oral cancer cells as well as primary cells for gene manipulation and expression in vivo. So far, several methods for a gene delivery have been performed to solve this problem. Magnetofection is one of the recent methods for gene transfer, and nanoparticles are applied under a magnetic field for DNA delivery. We investigated whether the magnetofection increases the efficiency of a gene delivery into several oral cell lines. By using a plasmid coding the green fluorescent protein (GFP), the efficiency of gene transfer by magnetofection was compared with those by using the calcium phosphate and the commercial transfection agent. Indeed, the magnetofection increased the green fluorescent signal in cells, suggested that this method apparently enhance the efficiency of gene delivery without any defects in various oral cancer cell lines. Finally, we have shown that magnetofection can be a useful technique for gene delivery to difficult-to-transfect cells to perform a functional study of genes in vivo.

Enhanced bone morphogenic protein adenoviral gene delivery to bone marrow stromal cells using magnetic nanoparticle

  • Lee, Jung-Tae;Jung, Jae-Whan;Choi, Jae-Yong;Kwon, Tae-Geon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.39 no.3
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    • pp.112-119
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    • 2013
  • Objectives: This study investigated the question of whether adenoviral magnetofection can be a suitable method for increasing the efficacy of gene delivery into bone marrow stromal cell (BMSC) and for generation of a high level of bone morphogenic protein (BMP) secretion at a minimized viral titer. Materials and Methods: Primary BMSCs were isolated from C57BL6 mice and transduced with adenoviral vectors encoding ${\beta}$ galactosidase or BMP2 and BMP7. The level of BMP secretion, activity of osteoblast differentiation, and cell viability of magnetofection were measured and compared with those of the control group. Results: The expression level of ${\beta}$ galactosidase showed that the cell transduction efficiency of AdLacZ increased according to the increased amount of magnetic nanoparticles. No change in cell viability was observed after magnetofection with 2 ${\mu}L$ of magnetic nanoparticle. Secretion of BMP2 or BMP7 was accelerated after transduction of AdBMP2 and 7 with magnetofection. AdBMP2 adenoviral magnetofection resulted in up to 7.2-fold higher secretion of BMP2, compared with conventional AdBMP2-transduced BMSCs. Magnetofection also induced a dramatic increase in secretion of BMP7 by up to 10-fold compared to the control. Use of only 1 multiplicity of infection (moi) of magnetofection with adenoviral transduction of AdBMP2 or AdBMP7 resulted in significantly higher transgene expression compared to 20 moi of conventional adenoviral transduction. Conclusion: Magnetic particle-mediated gene transudation is a highly efficient method of gene delivery to BMSCs. Magnetofection can lower the amount of viral particles while improving the efficacy of gene delivery.

Lipofectamine-2000 Assisted Magnetofection to Fibroblast Cells Using Polyethyleneimine-Fe3O4@SiO2 Nanoparticles

  • Jang, Eue-Soon;Park, Kyeong-Soon
    • Bulletin of the Korean Chemical Society
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    • v.33 no.8
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    • pp.2567-2573
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    • 2012
  • We successfully synthesized $Fe_3O_4@SiO_2$ nanoparticles with ultrathin silica layer of $1.0{\pm}0.5$ nm that polyethyleneimine (PEI) with low molecular weight of 2.0-4.0 kDa was covalently conjugated with the resulting $Fe_3O_4@SiO_2$ nanoparticles by silane coupling reaction. The PEI-$Fe_3O_4@SiO_2$ nanoparticles were further used as gene delivery vector for a human fibroblast cell (IMR-90) line. Gene transfection efficiency of the PEI-$Fe_3O_4@SiO_2$ complexes did not increase remarkably after magnetofection; however, the addition of Lipofectamine 2000 significantly increased the transfection efficiency of the PEI-$Fe_3O_4@SiO_2$ complexes. We believe that the present approach could be utilized for magnetofection as alternative to $Fe_3O_4$ nanoparticles conjugated with the PEI of high molecular weight thanks to its relatively low cytotoxicity and high transfection efficiency.

Comparison of Various Transfection Methods in Human and Bovine Cultured Cells

  • Jin, Longxun;Kim, Daehwan;Roh, Sangho
    • International Journal of Oral Biology
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    • v.39 no.4
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    • pp.177-185
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    • 2014
  • Transfection is a gene delivery tool that is a popular means of manipulating cellular properties, such as induced pluripotent stem cell (iPSC) generation by reprogramming factors (Yamanaka factors). However, the efficiency of transfection needs to be improved. In the present study, three transfection protocols - non-liposomal transfection (NLT), magnetofection and electroporation - were compared by analysis of their transfection efficiencies and cell viabilities using human dental pulp cells (hDPC) and bovine fetal fibroblasts (bFF) as cell sources. Enhanced green fluorescent protein gene was used as the delivery indicator. For magnetofection, Polymag reagent was administrated. NLT, FuGENE-HD and X-treme GENE 9 DNA transfection reagents were used for NLT. For electroporation, the $Neon^{TM}$ and $NEPA21^{TM}$ electroporators were tested. $Neon^{TM}$ electroporation showed highest transfection efficiency when compared with NLT, magnetofection, and $NEPA21^{TM}$ electroporation, with transfection efficiency of about 33% in hDPC and 50% in bFF, based on viable cell population in each cell type. These results suggest that transfection by $Neon^{TM}$ electroporation can be used to deliver foreign genes efficiently in human and bovine somatic cells.