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Magnet Assisted Transfection

To perform transfection with Magnetofection technology, you only need to buy the magnetic plate once and the reagents (magnetic particle formulations) appropriate to your needs. There is no further equipment required, so Magnetofection is an economical solution, especially for hard-to-transfect cells.

MagnetofectionTM is a novel, simple and highly efficient method to transfect cells In Vitro.

Two Basic types of ready-to-use Magnetofection™ reagents:

PolyMAG is a universally applicable magnetic particle preparation for high efficiency nucleic acid delivery. It is mixed in a one-step procedure with the nucleic acid to be transfected and has been used successfully with plasmid DNA, antisense oligonucleotides and siRNAs.

CombiMAG is a magnetic particle preparation designed to be combined with any commercially available transfection reagent such as polycations and lipids and can be associated with plasmid DNA, antisense oligonucleotides, siRNAs or viruses. It allows you to create your own magnetic gene vector based on your favourite transfection reagent.

 

 


     To perform transfection with Magnetofection technology, you only need to buy the magnetic plate once and the reagents
     (magnetic particle formulations) appropriate to your needs.

     There is no further equipment required, so Magnetofection is an economical solution, especially for hard-to-transfect cells.

     All Magnetofection starting kits from OZ Biosciences contain a magnetic plate and the reagents depending on your needs.
     It gives you a convenient solution to start your study. 

How to choose The right transfection reagent

 

     icon    PolyMag Neo: A reagent for all nucleic acids transfection

     icon    CombiMag: for enhancing all transfection reagents efficiency

     icon    ViroMag and ViroMag R/L: for virus transduction

     icon    AdenoMag: for adenovirus & AAV transduction NEW

     icon    Neuromag: Neurons specific transfection reagent

     icon    SilenceMag: for siRNA applications

     icon    LipoMag Kit : transfect all kind of cells with the best efficiency

An innovative transfection technology

Magnetic nanoparticles for nucleic acids and small molecules delivery

This method associates nucleic acids or other vectors with magnetic nanoparticles coated with cationic molecules. The magnetic nanoparticles are made of iron oxyde, which is fully biodegradable, coated with specific cationic proprietary molecules varying upon applications. Their association with the gene vectors (DNA, siRNA, ODN, virus, etc.) is achieved by salt-induced colloidal aggregation and electrostatic interaction.

Intracellular uptake of nanoparticles

The resulting molecular complexes are then targeted to and endocyted by cells, supported by an appropriate magnetic field. Membrane architecture and structure stay intact in contrast to other physical transfection methods that damage, create hole or electroshock the cell membranes. In addition the magnetic nanobeads are completely biodegradable and not toxic at the recommended doses and even higher. In this manner, the complete applied vector dose gets concentrated on the cells within a few minutes. Consequently, 100% of the cells get in contact with a significant vector dose.

Intracellular release of vectors and drugs         

The nucleic acids or other vectors are then released into the cytoplasm by different mechanisms depending on the formulation used. One is the proton sponge effect caused by cationic polymers coated on the nanoparticles that promotes endosome osmotic swelling, disruption of the endosomal membrane and intracellular release of associated molecules. Second is the destabilization of the endosome by cationic lipids coated on the particles that release the vectors into cells by flip-flop of cell negative lipids and charged neutralization. Third one is the usual viral mechanism when virus is used.

Inspired by the validated and recognized magnetic drug targeting technology, this innovative method is a revolution for transfection and infection.

 

MagnetoFection™ applications and results  

MagnetoFection™ has been successfully tested on a broad range of cells and cell lines. It is perfect for hard-to-transfect cells, primary cells and non dividing or slowly dividing cells. We have shown that combining magnetic nanoparticles to gene vectors of any kind results in a dramatic increase of uptake of these vectors and high transfection or delivery efficiency.

 

Primary Human Keratinocytes transfection  Primary Pig Chondrocytes transfection
   

 

siRNA delivery in HUVEC with SilenceMag   Adenovirus infection with ViroMag
   

 

MagnetoFection™ advantages compared to standard transfections

  • icon   Greatly improved transfection rates in terms of percentage of cells transfected compared to standard transfection. Very high transfection efficiencies with a wide range of cell types (many common cell lines as well as primary cells)
  • icon   successfully used with primary human cells and many other hard-to-transfect cells
  • icon   low cytotoxicity - Non toxic method with most commonly used cell lines

 

  • icon   Up to several thousand fold increased levels of transgene expression compared to standard transfections upon short-term incubation.
  • icon   functional with serum and serum-free
  • icon   Extremely short process time. A few minutes of incubation of cells with gene vectors are sufficient to generate high transfection efficiency, compared to several hours with standard procedures.
  • icon   highly cost-effective - no need for expensive instruments (like electroporator or particle gun)
  • icon   High transfection rates and transgene expression levels are achievable with extremely low vector doses, which allows to save expensive transfection reagents. 
  • icon   saves time and material.

 

MagnetoFection™ equipment

The only requirement for MagnetoFectionTM is a magnetic plate specifically designed for this application. The magnetic plate is a one-time buy and completely reusable, so you do not need expensive equipment contrary to approaches such as electroporation or gene gun. Two optimized magnetic plates with improved properties have been developed to allow you reaching the best levels of nanobeads delivery. Its design allows producing a magnetic field that magnetizes the nanoparticles in solution, forms a very strong gradient to attract them and covers all the surface of the plate. 

Super Magnetic Plate  Magnetic plate with 96 magnets Mega Magnetic plate
     
Convenient for all cell culture support
Catalog number # OZ_MF10000
 Adapted to 96-well plates
Catalog number #MF10096
To hold 4 culture dishes at one time #MF14000

 

MagnetoFection™ rapid and straightforward general protocol 

 

It is recommended to seed or plate the cells the day prior transfection. The suitable cell density will depend on the growth rate and the cells conditions. Cells should be 60-90% confluent at the time of Magnetofection (see the suggested cell number in the table below). For suspension cells, use the specific protocol given below. Immediately preceding transfection, the medium can be replaced with fresh medium (optionally without serum) if necessary.

Tissue Culture Dish Cell Number DNA Quantity (μg) Transfection Volume
96 well 0.5 – 2 x 10E4 0.1 – 0.5 0.2 mL
24 well 0.5 – 1 x 10E5 0.5 – 2.0 0.5 mL
12 well 1 – 2 x 10E5 2 – 4.0 1 mL
6 well 2 – 4 x 10E5 2 – 6.0 2 mL
60 mm dish 5 – 10 x 10E5 6 – 8.0 4 mL
90 - 100 mm dish 10 – 20 x 10E5 8 – 12.0 8 mL
T-75 flask 20 – 50 x 10E5 10 – 20.0 12 mL

The same protocol can be used to produce stably transfected cells except that 48 hours post transfection fresh medium containing the appropriate antibiotics are transferred to cells for selection. It is important to wait at least 48 hours before exposing the transfected cells to selection media.

Vectors are prepared in medium without serum and supplement or in physiological saline buffer. According to the standard Magnetofection protocol, the serum- and supplement-free vector cocktail is added to the cells that are cultured in complete medium. Therefore, the addition of the transfection solution will result in the dilution of standard culture medium. For most cell types, a medium change is not required after Magnetofection. However, it may be necessary for cells that are sensitive to serum/supplement concentration. Alternatively, the cells may be kept in serum-free medium during Magnetofection (up to 4 hours). In this case, a medium change will be required after Magnetofection.

Several protocol optimizations are available in the Appendix and upon request by contact fromular.

For more questions see FAQs

MEDIBENA Life Science & Diagnostic Solutions | Zeillergasse 38, 1170 Vienna, Austria, Europe | office@medibena.at | TEL: +43-699-1133-0065 | FAX: +43-1-2533-0339-577 | Skype: medibena | Register of companies: Handelsgericht Wien Österreich | FN: 392173a | VAT: ATU67954225 | EORI: ATEOS1000086649 | Dr. Monika Bencurova MEDIBENA Life Science E.U. | www.MEDIBENA.at