The effects of exposure of human neuroblastoma cells to a 50 Hz magnetic field on proliferation and differentiation in 2D and 3D cultures should be investigated.
The conventional 2D monolayer cultures were carried out by plating cells in 12-well plates. 3D cultures were created by using commercial 200-μm-thick polystyrene scaffolds.
Cells were divided into the following groups: 1) exposure to the magnetic field in 2D culture, 2) exposure to the magnetic field in 3D culture. For each exposure group, a separate sham exposure group was used. For proliferation experiments, cells were exposed for 72 hours. For dopaminergic differentiation experiments, cells were treated with retinoic acid for 3 days without MF exposure, followed by treatment wirth phorbol 12-myristate 13-acetate for further 3 days in combination with MF exposure.
| Exposure | Parameters |
|---|---|
|
Exposure 1:
50 Hz
Exposure duration:
72 hours
|
| Frequency | 50 Hz |
|---|---|
| Type | |
| Exposure duration | 72 hours |
| Exposure source | |
|---|---|
| Setup | exposure system consisted of two couples of square coils (two coils for each sub-system, arranged coaxially in Helmholtz configuration); the coil double wire configuration was used for sham exposure, which allowed to obtain a null magnetic field by using currents flowing in opposite directions; the magnetic field had a high homogeneity (95%) in the exposure volume of coils (20 × 20 × 10 cm3); temperature was kept at 37.0 ± 0.2 °C in both setups |
| Sham exposure | A sham exposure was conducted. |
| Measurand | Value | Type | Method | Mass | Remarks |
|---|---|---|---|---|---|
| magnetic flux density | 1 mT | effective value | measured and calculated | - | - |
Intracellular glutathione levels and SOD1 gene expression were significantly reduced in cells exposed to the magnetic field in 3D-cultures (group 2) compared to the sham exposure, whereas no change was observed in cells grown in 2D cultures (group 1). Moreover, a growth arrest and a significant increase in p21, TH, DAT and GAP43 gene expression was only found in group 2 compared to the sham exposure, indicating a synergy with the differentiating agents to stimulate neuroblastoma cell differentiation into a dopaminergic phenotype.
The authors concluded that exposure of human neuroblastoma cells to a 50 Hz magnetic field might change proliferation and differentiation in 3D cultures but not in 2D cultures.
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