Magnetofection-Based Transfection In Vitro: A Magnetic Field-Assisted Technique to Deliver Plasmids Into Primary Mouse Neuronal Cells Using Magnetic Nanoparticles
Published: April 30, 2023
Abstract
Source: Jacquier, A. et al. Modeling Charcot-Marie-Tooth Disease In Vitro by Transfecting Mouse Primary Motoneurons. J. Vis. Exp. (2019)
In this video, we demonstrate magnetically-guided transfection of plasmid DNA in primary neuronal cell culture. Magnetofection uses an external magnetic field to guide the delivery of plasmids bound to magnetic nanoparticles into cell cytoplasm.
Protocol
1. Poly-ornithine/Laminin (Po/L) Dish Coating NOTE: Volumes are adapted to coat a 24-well plate. If needed, put a sterile 12 mm coverslip in the well. Coat the wells with 500 µL of 10 µg/mL Poly-L-Ornithine solution dissolved in water. Let the wells settle at room temperature for 1 h. Remove the Poly-L-Ornithine solution and air-dry for 30 min. Coat the wells overnight at 37 °C with 500 µL…
Divulgazioni
The authors have nothing to disclose.
Materials
| Plasmid pCAGEN | Addgene | #11160 | |
| Neuron cell culture medium | ThermoFisher Scientific | A3582901 | Neurobasal Plus medium |
| Poly-DL-Ornithine | Sigma-Aldrich | P8638 | |
| Laminin | Sigma-Aldrich | L2020 | |
| Sodium bicarbonate | ThermoFisher Scientific | 25080094 | |
| L-15 medium | ThermoFisher Scientific | 11415056 | |
| PBS w/o Ca Mg | ThermoFisher Scientific | 14190144 | Without Mg2+ Ca2+ |
| Round coverslip | NeuVitro, Knittel glass | GG-12-Pre | 12 mm |
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Citazione di questo articolo
Magnetofection-Based Transfection In Vitro: A Magnetic Field-Assisted Technique to Deliver Plasmids Into Primary Mouse Neuronal Cells Using Magnetic Nanoparticles. J. Vis. Exp. (Pending Publication), e20997, doi: (2023).