Publication detail

DNA intracellular delivery into 3T3 cell line using fluorescence magnetic ferumoxide nanoparticles

SVOBODA, O. SKOPALÍK, J. BAIAZITOVA, L. ČMIEL, V. POTOČŇÁK, T. PROVAZNÍK, I. FOHLEROVÁ, Z. HUBÁLEK, J.

Original Title

DNA intracellular delivery into 3T3 cell line using fluorescence magnetic ferumoxide nanoparticles

English Title

DNA intracellular delivery into 3T3 cell line using fluorescence magnetic ferumoxide nanoparticles

Type

conference paper

Language

en

Original Abstract

Gene delivery is a widespread strategy in current experimental medicine. In this work, we report a method for low-toxic intracellular DNA vector delivery and post transfection localisation of this vector in mouse embryonic fibroblast cell lines. The surface of modified ferumoxide nanoparticles conjugated with Rhodamine B isothiocyanate (FeNV-Rh) was modified with linear polyethyleneimine and medium molecular weight chitosan to increase Accelerated Sensor of Action Potentials DNA vector adhesion. The size of the FeNV-Rh/DNA transfection complex was studied using dynamic light scattering (DLS) and scanning electron microscopy (SEM) techniques. The transfection complex internalisation of plasmid expression and FeNV-Rh, and stability of rhodamine fluorescence in intracellular space were observed at time periods 6, 12, 24 and 48 h post transfection. Results showed high transfection complex intracellular biocompatibility—cell viability after Rh-MNP labelling was higher than 97% 24 h after transfection, and higher than 95% after the next 24 h. Selective FeNV-Rh localisation in the lysosomes was quantified. More than 82% of nanoparticles were localised in the lysosomes 12 h post transfection and 94% of lysosomes had a significant and long-term deposit of nanoparticles. DNA vector expression was visible in >65% of the cells and precise protein localisation on the cell membrane was confirmed using confocal microscopy.

English abstract

Gene delivery is a widespread strategy in current experimental medicine. In this work, we report a method for low-toxic intracellular DNA vector delivery and post transfection localisation of this vector in mouse embryonic fibroblast cell lines. The surface of modified ferumoxide nanoparticles conjugated with Rhodamine B isothiocyanate (FeNV-Rh) was modified with linear polyethyleneimine and medium molecular weight chitosan to increase Accelerated Sensor of Action Potentials DNA vector adhesion. The size of the FeNV-Rh/DNA transfection complex was studied using dynamic light scattering (DLS) and scanning electron microscopy (SEM) techniques. The transfection complex internalisation of plasmid expression and FeNV-Rh, and stability of rhodamine fluorescence in intracellular space were observed at time periods 6, 12, 24 and 48 h post transfection. Results showed high transfection complex intracellular biocompatibility—cell viability after Rh-MNP labelling was higher than 97% 24 h after transfection, and higher than 95% after the next 24 h. Selective FeNV-Rh localisation in the lysosomes was quantified. More than 82% of nanoparticles were localised in the lysosomes 12 h post transfection and 94% of lysosomes had a significant and long-term deposit of nanoparticles. DNA vector expression was visible in >65% of the cells and precise protein localisation on the cell membrane was confirmed using confocal microscopy.

Keywords

DNA, delivery, 3T3 cells, fluorescence, ferumoxide magnetic nanoparticles, FeNV-Rh

Released

30.05.2018

Publisher

Springer

Location

Singapore

ISBN

978-981-10-9023-3

Book

World Congress on Medical Physics and Biomedical Engineering 2018

Pages from

149

Pages to

153

Pages count

5

URL

BibTex


@inproceedings{BUT148237,
  author="Ondřej {Svoboda} and Josef {Skopalík} and Larisa {Baiazitova} and Vratislav {Čmiel} and Tomáš {Potočňák} and Ivo {Provazník} and Zdenka {Fohlerová} and Jaromír {Hubálek}",
  title="DNA intracellular delivery into 3T3 cell line using fluorescence magnetic ferumoxide nanoparticles",
  annote="Gene delivery is a widespread strategy in current experimental medicine. In this work, we report a method
for low-toxic intracellular DNA vector delivery and post transfection localisation of this vector in mouse
embryonic fibroblast cell lines. The surface of modified ferumoxide nanoparticles conjugated with Rhodamine
B isothiocyanate (FeNV-Rh) was modified with linear polyethyleneimine and medium molecular
weight chitosan to increase Accelerated Sensor of Action Potentials DNA vector adhesion. The size of the
FeNV-Rh/DNA transfection complex was studied using dynamic light scattering (DLS) and scanning
electron microscopy (SEM) techniques. The transfection complex internalisation of plasmid expression
and FeNV-Rh, and stability of rhodamine fluorescence in intracellular space were observed at time periods
6, 12, 24 and 48 h post transfection. Results showed high transfection complex intracellular
biocompatibility—cell viability after Rh-MNP labelling was higher than 97% 24 h after transfection, and
higher than 95% after the next 24 h. Selective FeNV-Rh localisation in the lysosomes was quantified.
More than 82% of nanoparticles were localised in the lysosomes 12 h post transfection and 94% of
lysosomes had a significant and long-term deposit of nanoparticles. DNA vector expression was visible in
>65% of the cells and precise protein localisation on the cell membrane was confirmed using confocal
microscopy.",
  address="Springer",
  booktitle="World Congress on Medical Physics and Biomedical Engineering 2018",
  chapter="148237",
  doi="10.1007/978-981-10-9023-3_27",
  howpublished="online",
  institution="Springer",
  number="3",
  year="2018",
  month="may",
  pages="149--153",
  publisher="Springer",
  type="conference paper"
}