Exposure of the SH-SY5Y Human Neuroblastoma Cells to 50-Hz Magnetic Field: Comparison Between Two-Dimensional (2D) and T
- PDF / 1,403,533 Bytes
- 16 Pages / 595.276 x 790.866 pts Page_size
- 116 Downloads / 165 Views
ORIGINAL ARTICLE
Exposure of the SH-SY5Y Human Neuroblastoma Cells to 50-Hz Magnetic Field: Comparison Between Two-Dimensional (2D) and Three-Dimensional (3D) In Vitro Cultures Claudia Consales 1 & Alessio Butera 2 & Caterina Merla 1 & Emanuela Pasquali 1 & Vanni Lopresto 1 & Rosanna Pinto 1 & Maria Pierdomenico 1 & Mariateresa Mancuso 1 & Carmela Marino 1 & Barbara Benassi 1 Received: 29 January 2020 / Accepted: 30 October 2020 # The Author(s) 2020
Abstract We here characterize the response to the extremely low-frequency (ELF) magnetic field (MF, 50 Hz, 1 mT) of SH-SY5Y human neuroblastoma cells, cultured in a three-dimensional (3D) Alvetex® scaffold compared to conventional two-dimensional (2D) monolayers. We proved that the growing phenotype of proliferating SH-SY5Y cells is not affected by the culturing conditions, as morphology, cell cycle distribution, proliferation/differentiation gene expression of 3D-cultures overlap what reported in 2D plates. In response to 72-h exposure to 50-Hz MF, we demonstrated that no proliferation change and apoptosis activation occur in both 2D and 3D cultures. Consistently, no modulation of Ki67, MYCN, CCDN1, and Nestin, of invasiveness and neoangiogenesis-controlling genes (HIF-1α, VEGF, and PDGF) and of microRNA epigenetic signature (miR-21-5p, miR-222-3p and miR-133b) is driven by ELF exposure. Conversely, intracellular glutathione content and SOD1 expression are exclusively impaired in 3D-culture cells in response to the MF, whereas no change of such redox modulators is observed in SH-SY5Y cells if grown on 2D monolayers. Moreover, ELF-MF synergizes with the differentiating agents to stimulate neuroblastoma differentiation into a dopaminergic (DA) phenotype in the 3D-scaffold culture only, as growth arrest and induction of p21, TH, DAT, and GAP43 are reported in ELF-exposed SH-SY5Y cells exclusively if grown on 3D scaffolds. As overall, our findings prove that 3D culture is a more reliable experimental model for studying SH-SY5Y response to ELF-MF if compared to 2D conventional monolayer, and put the bases for promoting 3D systems in future studies addressing the interaction between electromagnetic fields and biological systems. Keywords Extremely low-frequency magnetic field . SH-SY5Y . In vitro . 3D culture . MicroRNAs
Abbreviations 2D Two-dimensional 3D Three-dimensional AD Alzheimer’s’ disease ALS Amyotrophic lateral sclerosis B-field Induction magnetic field E-field Electric field ELF-MF Extremely low-frequency magnetic field
* Barbara Benassi [email protected] 1
2
Division of Health Protection Technologies, ENEA-Casaccia Italian National Agency for New Technologies, Energy and Sustainable Economic Development, Via Anguillarese 301, 00123 Rome, Italy Experimental Medicine and Surgery, University of Rome Tor Vergata, 00133 Rome, Italy
H&E HIC IF J MF PMA RA RMS WST-1
Hematoxylin and eosin Immunohistochemistry Immunofluorescence Current density Magnetic field Phorbol 12-myristate 13-acetate Retinoic acid Root mean square Water Soluble Tetrazolium Salt-1
Int
Data Loading...
