A Novel 3D Scaffold for Cell Growth to Asses Electroporation Efficacy

Abstract

Tumor electroporation (EP) refers to the permeabilization of the cell membrane by means of short electric pulses thus allowing the potentiation of chemotherapeutic drugs. Standard plate adhesion 2D cell cultures can simulate the in vivo environment only partially due to lack of cell–cell interaction and extracellular matrix (ECM). In this study, we assessed a novel 3D scaffold for cell cultures based on hyaluronic acid and ionic-complementary self-assembling peptides (SAPs), by studying the growth patterns of two different breast carcinoma cell lines (HCC1569 and MDA-MB231). This 3D scaffold modulates cell shape and induces extracellular matrix deposit around cells. In the MDA-MB 231 cell line, it allows three-dimensional growth of structures known as spheroids, while in HCC1569 it achieves a cell organization similar to that observed in vivo. Interestingly, we were able to visualize the electroporation effect on the cells seeded in the new scaffold by means of standard propidium iodide assay and fluorescence microscopy. Thanks to the presence of cell–cell and cell–ECM interactions, the new 3D scaffold may represent a more reliable support for EP studies than 2D cancer cell cultures and may be used to test new EP-delivered drugs and novel EP protocols.

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

“…Moreover, the 3D scaffold of hyaluronic acid and SAPs induces a consistent production and deposit of ECM components around cells, thus reproducing a common feature of several human breast malignancies that show an abundant fibrous component, with collagen constituent, and scarce cellular component [38,56,57]. We showed that, as further demonstrated in [50], our 3D model can represent a suitable experimental platform to test GET protocols on mammary tumor cells surrounded by ECM.…”

“…We previously reported on EP experiments on breast cancer cells seeded in such a collagen-free 3D scaffold [50]. Thanks to the presence of cell-cell and cell-ECM interactions, the new 3D scaffold may represent more reliable support for EP studies than 2D cancer cell cultures, and it may be used to test new EP-delivered drugs and novel EP protocols [50].…”

“…HCC1954 and MDA-MB231 cell lines were grown in the new collagen-free 3D scaffold and underwent the application of a GET protocol consisting of EP-based transfection of the plasmid pEGFP-C3 using linear electrodes (8 voltage pulses, 1300 V/cm, 100 µs long at 1 Hz). These electrical parameters were previously demonstrated to assure a high EP efficiency in cells seeded in monolayer or the same 3D scaffold [50]. To verify the reliability of the novel 3D scaffold as an ex vivo model for GET protocols, we applied EP through linear electrodes on HCC1954 and MDA-MB231 3D cell cultures.…”

“…We previously demonstrated that it could reliably reproduce in vivo tissue architecture [48,49]. This scaffold result was particularly suitable to reproduce a tumor microenvironment where cells are embedded in a myxoid stroma, characterized by abundant amounts of hyaluronic acid and proteoglycans, low collagen, and elastin content [50].…”

<p>Prediction and Monitoring Method for Breast Cancer: A Case Study for Data from the University Hospital Centre of Coimbra</p>

“…Robust predictive models based on data that may be collected in routine consultations and blood analysis are sought to provide an important contribution by offering more screening tools, and are important for detecting whether there is a change in the breast cancer index. 2,3 Statistical process control (SPC) has been frequently used for fault detection. [4][5][6] One major concern of SPC is whether there has been a change of distribution from the target in the process, that is, the process has gone out of control.…”