Electroporation of cells is usually studied using cell suspensions or monolayer cultures. 3D scaffolds for cell culture have been recently designed in order to reproduce in vitro the complex and multifactorial environment experimented in vivo by cells. In fact, it is well known that 2D cell cultures are not able to simulate the complex interactions between the cells and their extracellular matrix (ECM). Recently, some examples of 3D models, like spheroids, have been investigated also in the electroporation field. Spheroids have been proposed in electrochemotherapy (ECT) studies to mimic tumor in vivo conditions: they are easy-to-handle 3D models but their sensitivity to electric field pulses depends from their diameter and, more interestingly, despite being relevant for intercellular junctions, they are not so much so for cell-ECM interactions.

In this work, we propose a 3D macroscopic myxoid matrix for cell culture that would mimic the in vivo environment of myxoid stroma tumors. The myxoid stroma consists of abundant basic substances with large amounts of glycosaminoglycans (hyaluronic acid) and proteoglycans, poor collagen fibers and no elastin content. In the proposed approach, tumor cells seeded on 3D scaffolds mimic of myxoid stroma can establish both cell-cell and cell-ECM 3D interactions.

The MCF7 cells (human breast adenocarcinoma cell line) were seeded in complete culture medium. Cell cultures were incubated at 37 °C for either 24 h, 3 days or 7 day. Some samples were used to assess cell vitality using 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) test and others for electroporation tests and for histopathological analysis. The electroporation has been verified by the fluorescent dye Propidium cellular uptake.

The proposed myxoid stroma scaffold induces cell proliferation and shows fibrous structures produced by cells, the concentration of which increases with culture time. The proposed matrix will be used for further investigations as a new scaffold for cell culture. Tumor cells grown into these new scaffolds will be used to evaluate electroporation including the stroma effect.

通常使用细胞悬浮液或单层培养物研究细胞的电穿孔。最近已设计出用于细胞培养的3D支架，以在体外复制细胞体内实验的复杂多因素环境。实际上，众所周知2D细胞培养不能模拟细胞与其细胞外基质（ECM）之间的复杂相互作用。最近，还在电穿孔领域研究了3D模型的一些示例，例如球体。在电化学疗法（ECT）研究中已提出球状体来模拟体内肿瘤：它们是易于处理的3D模型，但它们对电场脉冲的敏感性取决于其直径，更有趣的是，尽管它们与细胞间连接有关，但它们对于细胞-ECM交互作用则不那么重要。

在这项工作中，我们提出了一种用于细胞培养的3D宏观类胶质基质，该基质将模仿类胶质基质肿瘤的体内环境。粘液样基质由丰富的碱性物质组成，具有大量的糖胺聚糖（透明质酸）和蛋白聚糖，胶原蛋白纤维较弱，且不含弹性蛋白。在提出的方法中，接种在粘液样基质的3D支架模拟物上的肿瘤细胞可以建立细胞-细胞和细胞-ECM 3D相互作用。

将MCF7细胞（人乳腺癌细胞系）接种在完全培养基中。将细胞培养物在37°C下孵育24小时，3天或7天。一些样品用于通过3-（4,5-二甲基噻唑基-2）-2,5-二苯基四唑溴化物（MTT）测试来评估细胞活力，而其他样品则用于电穿孔测试和组织病理学分析。电穿孔已被荧光染料丙啶细胞摄取所证实。

拟议的粘液样基质支架诱导细胞增殖并显示出细胞产生的纤维结构，其浓度随培养时间而增加。拟议的基质将用作细胞培养的新支架用于进一步的研究。生长在这些新支架中的肿瘤细胞将用于评估电穿孔，包括基质效应。