Publication detail

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

SVOBODA, O. FOHLEROVÁ, Z. BAIAZITOVA, L. MLÝNEK, P. SAMOUYLOV, K. PROVAZNÍK, I. HUBÁLEK, J.

Original Title

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

English Title

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

Type

journal article in Web of Science

Language

en

Original Abstract

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.

English abstract

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.

Keywords

Polyethyleneimine, magnetic nanoparticles, transfection, patch clamp, cytotoxicity, ASAP1

Released

01.08.2018

Publisher

American Scientific Publishers

Location

Valencia, California, USA

Pages from

1505

Pages to

1514

Pages count

10

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"
}