Detail publikace

Transfection by Polyethyleneimine-coated Magnetic Nanoparticles: Fine-tuning the Condition for Electrophysiological Experiments

Originální název

Transfection by Polyethyleneimine-coated Magnetic Nanoparticles: Fine-tuning the Condition for Electrophysiological Experiments

Anglický název

Transfection by Polyethyleneimine-coated Magnetic Nanoparticles: Fine-tuning the Condition for Electrophysiological Experiments

Jazyk

en

Originální abstrakt

A non-viral tool for the delivery of nucleic acids termed magnetofection was recently developed as a promising transgenic technique with high transfection efficiency for gene delivery into mammalian cells. Despite the fact that transfection efficiency was the objective in the past, the post-transfection cell morphology and the essential gigaseal formation between cells and patch clamp glass electrodes have not been studied in detail. The cell viability and fluorescent response of Accelerated Sensor of Action Potentials (ASAP1) were studied in somatic HEK293 cells with respect to preserving physiological cell behavior and morphology. The DNA vector (pcDNA3.1/Puro-CAG-ASAP1) was intracellularly delivered by DNA/polyethyleneimine/magnetic nanoparticles and the transfection protocols varied in complex formations were optimized with respect to transfection rate, cytotoxicity of modified nanoparticles and essential gigaseal formation needed for patch clamp technique. A patch clamp study of transfected cells was carried out 72 hours post-transfection. Our results showed the best complex formation in order DNA/magnetic nanoparticle/polyethyleneimine that provides 51.82% transfection efficiency, 83.45% of patch clamp applicable cells, and 90.15% of gigasealed patch clamp applicable cells. A significant difference in fluorescent response of transfected cells was not found compared to control. Thus, these observations suggested that a large amount of the cells were able to create a gigaseal with a glass electrode 72 hours from transfection despite the lower transfection efficiencies.

Anglický abstrakt

A non-viral tool for the delivery of nucleic acids termed magnetofection was recently developed as a promising transgenic technique with high transfection efficiency for gene delivery into mammalian cells. Despite the fact that transfection efficiency was the objective in the past, the post-transfection cell morphology and the essential gigaseal formation between cells and patch clamp glass electrodes have not been studied in detail. The cell viability and fluorescent response of Accelerated Sensor of Action Potentials (ASAP1) were studied in somatic HEK293 cells with respect to preserving physiological cell behavior and morphology. The DNA vector (pcDNA3.1/Puro-CAG-ASAP1) was intracellularly delivered by DNA/polyethyleneimine/magnetic nanoparticles and the transfection protocols varied in complex formations were optimized with respect to transfection rate, cytotoxicity of modified nanoparticles and essential gigaseal formation needed for patch clamp technique. A patch clamp study of transfected cells was carried out 72 hours post-transfection. Our results showed the best complex formation in order DNA/magnetic nanoparticle/polyethyleneimine that provides 51.82% transfection efficiency, 83.45% of patch clamp applicable cells, and 90.15% of gigasealed patch clamp applicable cells. A significant difference in fluorescent response of transfected cells was not found compared to control. Thus, these observations suggested that a large amount of the cells were able to create a gigaseal with a glass electrode 72 hours from transfection despite the lower transfection efficiencies.

BibTex


@article{BUT146879,
  author="Ondřej {Svoboda} and Zdenka {Fohlerová} and Larisa {Baiazitova} and Petr {Mlýnek} and Konstantin {Samouylov} and Ivo {Provazník} and Jaromír {Hubálek}",
  title="Transfection by Polyethyleneimine-coated Magnetic Nanoparticles: Fine-tuning the Condition for Electrophysiological Experiments",
  annote="A non-viral tool for the delivery of nucleic acids termed magnetofection was recently developed as a promising transgenic technique with high transfection efficiency for gene delivery into mammalian cells. Despite the fact that transfection efficiency was the objective in the past, the post-transfection cell morphology and the essential gigaseal formation between cells and patch clamp glass electrodes have not been studied in detail. The cell viability and fluorescent response of Accelerated Sensor of Action Potentials (ASAP1) were studied in somatic HEK293 cells with respect to preserving physiological cell behavior and morphology. The DNA vector (pcDNA3.1/Puro-CAG-ASAP1) was intracellularly delivered by DNA/polyethyleneimine/magnetic nanoparticles and the transfection protocols varied in complex formations were optimized with respect to transfection rate, cytotoxicity of modified nanoparticles and essential gigaseal formation needed for patch clamp technique. A patch clamp study of transfected cells was carried out 72 hours post-transfection. Our results showed the best complex formation in order DNA/magnetic nanoparticle/polyethyleneimine that provides 51.82% transfection efficiency, 83.45% of patch clamp applicable cells, and 90.15% of gigasealed patch clamp applicable cells. A significant difference in fluorescent response of transfected cells was not found compared to control. Thus, these observations suggested that a large amount of the cells were able to create a gigaseal with a glass electrode 72 hours from transfection despite the lower transfection efficiencies.",
  address="American Scientific Publishers",
  chapter="146879",
  doi="10.1166/jbn.2018.2602",
  howpublished="online",
  institution="American Scientific Publishers",
  number="8",
  volume="14",
  year="2018",
  month="august",
  pages="1505--1514",
  publisher="American Scientific Publishers",
  type="journal article in Web of Science"
}