基于有限元建模的BT-20和SKOV-3细胞系电穿孔方案的建立

C. E. V. Tizatl, A. L. V. Tizatl, J. Osorio-Trujillo, P. Talamás-Rohana, S. Rodriguez-Cuevas, L. Leija, A. Vera
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引用次数: 1

摘要

目前皮肤和皮下肿瘤的治疗方法是基于电穿孔的应用,即电疗(ECT)。我们推断,从这些恶性肿瘤报道的结果,ECT可能用于根除原发性深部乳腺恶性肿瘤。因此,本研究的目的是基于BT-20细胞电穿孔的有限元模型和体外电穿孔,建立一种针对乳腺组织的电穿孔方案。将这些电穿孔方案与第二种细胞系SKOV-3(一种卵巢癌来源的细胞系)获得的电穿孔方案进行比较,以确定细胞来源是否对模拟体外条件的计算模型预测的电穿孔方案的效率有影响。通过荧光显微镜和MTT法测定碘化丙啶的摄取和细胞活力,验证了其有效性。结果表明,在8个150 V脉冲、脉冲宽度100 μs、脉冲重复频率1 s的条件下,SK-OV-3细胞株的细胞存活率为98.7%,与计算模型预测相符。但是,要使BT-20细胞株的细胞存活率达到89.1%的最大值,必须将该方案改进为8个140 V脉冲,脉冲宽度为100 μs,脉冲重复频率为1 s。本研究方法的一个局限性是不典型地使用悬浮的SK-OV-3和BT-20细胞,因为它们是贴壁的,也就是说,这些细胞需要基质来保持其上皮和形态特征,这可能导致是否对贴壁细胞进行电穿孔。尽管如此,这项研究的贡献是确定了人类乳腺癌细胞系可逆电穿孔的第一个特定方案。因此,鼓励在实验应用之前计划建立器官特异性电穿孔协议。
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Establishment of Electroporation Protocols in BT-20 and SKOV-3 Cell Lines based on Finite Element Modeling
A current treatment method of cutaneous and subcutaneous tumors is based on an application of electroporation, known as Electrochemotherapy (ECT). We infer, from the outcomes reported on these malignancies, that ECT might be utilized for eradication of primary deep-seated breast malignancies. The aim of this work is therefore, to establish an electroporation protocol specific for breast tissue, based on a finite element model of electroporation of BT-20 cells, and in vitro electroporation. These electroporation protocols were compared with those obtained with a second cell line SKOV-3, an ovarian carcinoma derived cell line, in order to determine whether the cellular origin has an effect on efficiency of an electroporation protocol predicted by computational models simulating in vitro conditions. Efficiency was verified through the determination of propidium iodide uptake and cell viability by epifluorescence microscopy and MTT assay. The results show that a protocol of 8 pulses of 150 V, pulse width of 100 μs, and pulse repetition frequency of 1 s, leading to a cell viability of 98.7 % in SK-OV-3 cell line is in accordance with the computational model predictions. Nonetheless, this protocol must be modified to a protocol of 8 pulses of 140 V, pulse width of 100 μs, and pulse repetition frequency of 1 s, so that a maximal cell viability of 89.1 % in BT-20 cell line can be obtained. A limitation of the methodology in this work is the atypical use of suspended SK-OV-3 and BT-20 cell, since they are adherent i.e., these cells require a substratum to keep their epithelial and morphological features which may lead to significant variations whether electroporation of attached cells was carried out. Nevertheless, the contribution of this research is the determination of a first specific protocol of reversible electroporation for a human breast cancer cell line. Consequently, the establishment of organ-specific electroporation protocols planned prior to experimental application is encouraged.
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