Magnetically guided lentiviral‐mediated transduction of airway epithelial cells

Abstract

We conclude that magnetofection can facilitate in vitro LV-mediated transduction of airway epithelial cells, in the absence of overt cytotoxicity and maintaining epithelial integrity, by lowering the necessary vector dose and reducing the incubation time required to achieve efficient transduction.

Magnetofection Enhances Lentiviral-Mediated Transduction of Airway Epithelial Cells through Extracellular and Cellular Barriers

“…Similar to what we found with another airway epithelial cell line (i.e., 16HBE bronchial cells [13]), the transduction in the absence of a magnetic field led to some enhancement, which was not significant. However, high transduction levels can be associated with increased cytotoxicity.…”

“…With R/L magnetoparticles, the enhancement effect was up to 2.1-fold. As noticed before with other respiratory epithelial cell lines [13], even in the absence of magnetic forces, the highest dose of magnetoparticles used (12 μL) was able to give an enhancement effect of 2.8- and 2.5-fold with VM and R/L, respectively. The following experiments were carried out with lentiviral magnetoparticles obtained with 2 μL of VM, since 12 μL had resulted in enhanced cytotoxicity in our previous study [13].…”

“…Magnetofection has been used in the field of respiratory epithelial cells to increase nonviral-mediated gene transfer [18,19,20,21] and to study the mechanism of internalization [22]. Although many studies have demonstrated that by using viral magnetofection systems, the expression of a transgene is often significantly higher in comparison to only inoculating with virus alone in many cell types [23,24,25,26,27], our previous work was the first to show that LV-mediated transduction of bronchial epithelial cells is enhanced by magnetofection [13]. Moreover, only a few studies have demonstrated the usefulness of magnetofection in targeting applications that require penetration of a cellular or a tessutal barrier [28].…”

“…After different time points, cells were washed two times with PBS and, subsequently, complete medium was added. Cells were incubated at 37 °C 5%CO 2 for 72 h, and then analyzed by flow cytometry (FACScan apparatus, Becton-Dickinson, San Jose, CA, USA) to evaluate efficiency, as previously described [13]. …”

“…Thus, the local confinement of nucleic acid delivery is an important requirement for gene delivery to the airway epithelium. Previously, we have shown that magnetofection can decrease the dose and the time of a lentiviral vector (LV) incubation with airway epithelial cells while maintaining a meaningful efficiency [13]. Furthermore, magnetofection resulted in at least 20-fold higher gene transfer levels than LV alone in a polarized model of airway epithelium.…”