Cell morphology impact on the set-up of electroporation protocols for in-suspension and adhered breast cancer cells.

In order to establish cancer-type-specific electroporation protocols for breast cancer, electroporation was performed in two modalities: in-suspension and adhered cells. Electroporation of cell suspensions was carried out through commercial electroporation cuvettes whereas a novel electrode for electroporation of adhered cells was designed and manufactured aimed to preserve cell structure, to provide a closer model to an scenario, and as a means to visualize the mechanical effects of electroporation on the cell membrane by using scanning electron microscopy. Electroporation protocols and electric field thresholds were predicted and experimentally tuned through propidium iodide uptake and cell viability. Three breast-cancer cell lines (BT-20, MCF-7 and HCC1419) and a non-cancerous cell line (BEAS-2B) were used. Cancerous cells responded differently to electroporation depending on the electric parameters, cell histology, the cell culture modality, and the cell morphology (membrane thickness mainly), which was evaluated trough confocal and transmission electron microscopy. Particularly, it was found that electrochemotherapy may represent a promising alternative as an adjuvant treatment of metastatic breast tumours, and as a neoadjuvant therapy for Her2/neu tumours. Oppositely, triple negative breast tumours may show a high sensitivity to electroporation and therefore, they could be efficiently treated with irreversible electroporation. On the other hand, noncancerous cells demanded the highest voltage in both cell culture modalities in order to be electroporated. Hence, these cells in suspension may provide a reliable, easy-to-perform, low-cost model for the development of electroporation protocols for eradication of healthy tissue around a tumour in a safety margin.