C. E. V. Tizatl, A. L. V. Tizatl, J. Osorio-Trujillo, P. Talamás-Rohana, S. Rodriguez-Cuevas, L. Leija, A. Vera
{"title":"基于有限元建模的BT-20和SKOV-3细胞系电穿孔方案的建立","authors":"C. E. V. Tizatl, A. L. V. Tizatl, J. Osorio-Trujillo, P. Talamás-Rohana, S. Rodriguez-Cuevas, L. Leija, A. Vera","doi":"10.1109/ICEEE.2018.8533974","DOIUrl":null,"url":null,"abstract":"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.","PeriodicalId":6924,"journal":{"name":"2018 15th International Conference on Electrical Engineering, Computing Science and Automatic Control (CCE)","volume":"21 1","pages":"1-5"},"PeriodicalIF":0.0000,"publicationDate":"2018-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":"{\"title\":\"Establishment of Electroporation Protocols in BT-20 and SKOV-3 Cell Lines based on Finite Element Modeling\",\"authors\":\"C. E. V. Tizatl, A. L. V. Tizatl, J. Osorio-Trujillo, P. Talamás-Rohana, S. Rodriguez-Cuevas, L. Leija, A. Vera\",\"doi\":\"10.1109/ICEEE.2018.8533974\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"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.\",\"PeriodicalId\":6924,\"journal\":{\"name\":\"2018 15th International Conference on Electrical Engineering, Computing Science and Automatic Control (CCE)\",\"volume\":\"21 1\",\"pages\":\"1-5\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2018-09-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"2018 15th International Conference on Electrical Engineering, Computing Science and Automatic Control (CCE)\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1109/ICEEE.2018.8533974\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"2018 15th International Conference on Electrical Engineering, Computing Science and Automatic Control (CCE)","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1109/ICEEE.2018.8533974","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
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.