Transfection by polyethyleneimine-coated magnetic nanoparticles: Fine-tuning the condition for electrophysiological experiments

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. © 2018 American Scientific Publishers.

Авторы
Svoboda O.1, 2 , Fohlerova Z.2, 3, 4 , Baiazitova L.1 , Mlynek P. 5 , Samouylov K. 6 , Provaznik I.1 , Hubalek J.2, 3, 4
Издательство
American Scientific Publishers
Номер выпуска
8
Язык
Английский
Страницы
1505-1514
Статус
Опубликовано
Том
14
Год
2018
Организации
  • 1 Department of Biomedical Engineering, Faculty of Electrical Engineering and Communication, Brno University of Technology, Technicka 12, Brno, 616 00, Czech Republic
  • 2 Central European Institute of Technology, Brno University of Technology, Purkynova 123, Brno, 612 00, Czech Republic
  • 3 Department of Microelectronics, Faculty of Electrical Engineering and Communication, Brno University of Technology, Technicka 10, Brno, 616 00, Czech Republic
  • 4 SIX Centre, Department of Microelectronics, Faculty of Electrical Engineering and Communication, Brno University of Technology, Technicka 3058/10, Brno, 616 00, Czech Republic
  • 5 Department of Telecommunications, Faculty of Electrical Engineering and Communication, Brno University of Technology, Technicka 10, Brno, 616 00, Czech Republic
  • 6 Peoples' Friendship University of Russia, RUDN University, 6 Miklukho-Maklaya St, Moscow, 117198, Russian Federation
Ключевые слова
ASAP1; Cytotoxicity; Magnetic nanoparticles; Patch clamp; Polyethyleneimine; Transfection
Дата создания
19.10.2018
Дата изменения
19.10.2018
Постоянная ссылка
https://repository.rudn.ru/ru/records/article/record/6497/