Next Article in Journal
Identification of Dietary Pattern Networks Associated with Gastric Cancer Using Gaussian Graphical Models: A Case-Control Study
Next Article in Special Issue
Cytoskeletal Disruption after Electroporation and Its Significance to Pulsed Electric Field Therapies
Previous Article in Journal
DNA Damage/Repair Management in Cancers
Previous Article in Special Issue
New Deployable Expandable Electrodes in the Electroporation Treatment in a Pig Model: A Feasibility and Usability Preliminary Study
Article

The Efficiency of Gene Electrotransfer in Breast-Cancer Cell Lines Cultured on a Novel Collagen-Free 3D Scaffold

1
Department of Theoretical and Applied Sciences, University of Insubria, 21100 Varese, Italy
2
Department of Industrial Engineering, University of Padova, 35131 Padova, Italy
3
CNR-Institute of Biomembrane, Bioenergetics and Molecular Biotechnology, 70126 Bari, Italy
4
Department of Bioscience, Biotechnology and Biopharmaceutics, University of Bari, 70126 Bari, Itay
5
Department of Pharmaceutical and Pharmacological Sciences, University of Padova, 35131 Padova, Italy
6
Campus Bio-Medico University of Rome, 00128 Roma, Italy
7
Department of Surgical Oncological and Gastroenterological Sciences DISCOG, University of Padova, 35124 Padova, Italy
8
CNR-Institute of Translational Pharmacology, 00133 Roma, Italy
*
Authors to whom correspondence should be addressed.
Cancers 2020, 12(4), 1043; https://doi.org/10.3390/cancers12041043
Received: 28 February 2020 / Revised: 8 April 2020 / Accepted: 21 April 2020 / Published: 23 April 2020
Gene Electro-Transfer (GET) is a powerful method of DNA delivery with great potential for medical applications. Although GET has been extensively studied in vitro and in vivo, the optimal parameters remain controversial. 2D cell cultures have been widely used to investigate GET protocols, but have intrinsic limitations, whereas 3D cultures may represent a more reliable model thanks to the capacity of reproducing the tumor architecture. Here we applied two GET protocols, using a plate or linear electrode, on 3D-cultured HCC1954 and MDA-MB231 breast cancer cell lines grown on a novel collagen-free 3D scaffold and compared results with conventional 2D cultures. To evaluate the electrotransfer efficiency, we used the plasmid pEGFP-C3 encoding the enhanced green fluorescent protein (EGFP) reporter gene. The novel 3D scaffold promoted extracellular matrix deposition, which particularly influences cell behavior in both in vitro cell cultures and in vivo tumor tissue. While the transfection efficiency was similar in the 2D-cultures, we observed significant differences in the 3D-model. The transfection efficiency in the 3D vs 2D model was 44% versus 15% (p < 0.01) and 24% versus 17% (p < 0.01) in HCC1954 and MDA-MB231 cell cultures, respectively. These findings suggest that the novel 3D scaffold allows reproducing, at least partially, the peculiar morphology of the original tumor tissues, thus allowing us to detect meaningful differences between the two cell lines. Following GET with plate electrodes, cell viability was higher in 3D-cultured HCC1954 (66%) and MDA-MB231 (96%) cell lines compared to their 2D counterpart (53% and 63%, respectively, p < 0.001). Based on these results, we propose the novel 3D scaffold as a reliable support for the preparation of cell cultures in GET studies. It may increase the reliability of in vitro assays and allow the optimization of GET parameters of in vivo protocols. View Full-Text
Keywords: Gene Electro-Transfer (GET); electroporation; 3D cell cultures; scaffold; breast cancer Gene Electro-Transfer (GET); electroporation; 3D cell cultures; scaffold; breast cancer
Show Figures

Figure 1

MDPI and ACS Style

Sieni, E.; Dettin, M.; De Robertis, M.; Bazzolo, B.; Conconi, M.T.; Zamuner, A.; Marino, R.; Keller, F.; Campana, L.G.; Signori, E. The Efficiency of Gene Electrotransfer in Breast-Cancer Cell Lines Cultured on a Novel Collagen-Free 3D Scaffold. Cancers 2020, 12, 1043. https://doi.org/10.3390/cancers12041043

AMA Style

Sieni E, Dettin M, De Robertis M, Bazzolo B, Conconi MT, Zamuner A, Marino R, Keller F, Campana LG, Signori E. The Efficiency of Gene Electrotransfer in Breast-Cancer Cell Lines Cultured on a Novel Collagen-Free 3D Scaffold. Cancers. 2020; 12(4):1043. https://doi.org/10.3390/cancers12041043

Chicago/Turabian Style

Sieni, Elisabetta; Dettin, Monica; De Robertis, Mariangela; Bazzolo, Bianca; Conconi, Maria T.; Zamuner, Annj; Marino, Ramona; Keller, Flavio; Campana, Luca G.; Signori, Emanuela. 2020. "The Efficiency of Gene Electrotransfer in Breast-Cancer Cell Lines Cultured on a Novel Collagen-Free 3D Scaffold" Cancers 12, no. 4: 1043. https://doi.org/10.3390/cancers12041043

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Search more from Scilit
 
Search
Back to TopTop