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电穿孔法转染 Ishikawa 细胞:流式细胞术分析。

Electroporation of Ishikawa cells: analysis by flow cytometry.

机构信息

Department of Engineering and Architectural Studies, Ara Institute of Canterbury, Christchurch, New Zealand.

Department of Electrical and Computer Engineering, University of Canterbury, Christchurch, New Zealand.

出版信息

IET Nanobiotechnol. 2019 Feb;13(1):58-65. doi: 10.1049/iet-nbt.2018.5194.

Abstract

Electroporation facilitates loading of cells with molecules and substances that are normally membrane impermeable. Flow cytometry is used in this study to examine the effects of the application of electroporation-level monopolar electric field pulses of varying electrical field strength on Ishikawa endometrial adenocarcinoma cells. Analysis of the fluorescence versus forward scatter plots corroborates the well-recognised threshold and cell size dependence characteristics of electroporation, but also shows the progression of cell lysis and generation of particulate material. Two 500 µs monopolar rectangular pulses ranging from 1.0 × 10 to 2.5 × 10 V/m were used to electroporate the cells. Electroporation yields (fraction of viable cells exhibiting significant propidium iodide uptake) ranged from 0 to 97%, with viability ranging between 78 and 34% over the electric field strength range tested. The higher electric field strength pulses not only reduced cell viability, but also generated a substantial amount of sub-cellular sized particulate material indicating cells have been physically disrupted enough to create these particles.

摘要

电穿孔促进了通常对细胞膜不可渗透的分子和物质向细胞内的装载。本研究采用流式细胞术检测不同电场强度的电穿孔级单极电场脉冲对 Ishikawa 子宫内膜腺癌细胞的影响。荧光与前向散射图的分析证实了电穿孔的公认阈值和细胞大小依赖性特征,但也显示了细胞裂解和颗粒物质生成的进展。使用两个持续时间为 500µs 的单极矩形脉冲,范围从 1.0×10 到 2.5×10V/m,对细胞进行电穿孔。电穿孔产率(表现出碘化丙啶摄取显著增加的活细胞分数)从 0 到 97%不等,在测试的电场强度范围内,细胞活力在 78%到 34%之间。较高的电场强度脉冲不仅降低了细胞活力,而且产生了大量亚细胞大小的颗粒物质,表明细胞已经被物理破坏到足以产生这些颗粒的程度。

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本文引用的文献

1
Electroporation for nanomedicine: a review.
J Mater Chem B. 2017 Apr 21;5(15):2726-2738. doi: 10.1039/c7tb00038c. Epub 2017 Mar 15.
3
A review of basic to clinical studies of irreversible electroporation therapy.
IEEE Trans Biomed Eng. 2015 Jan;62(1):4-20. doi: 10.1109/TBME.2014.2367543.
4
Scaling relationship and optimization of double-pulse electroporation.
Biophys J. 2014 Feb 18;106(4):801-12. doi: 10.1016/j.bpj.2013.12.045.
5
Assessment of cell viability.
Curr Protoc Cytom. 2013;Chapter 9:9.2.1-9.2.26. doi: 10.1002/0471142956.cy0902s64.
6
Electro-gene-transfer as a new tool for cancer immunotherapy in animals.
Vet Comp Oncol. 2014 Dec;12(4):310-8. doi: 10.1111/vco.12006. Epub 2012 Oct 25.
7
A brief overview of electroporation pulse strength-duration space: a region where additional intracellular effects are expected.
Bioelectrochemistry. 2012 Oct;87:236-43. doi: 10.1016/j.bioelechem.2012.02.007. Epub 2012 Mar 14.
8
Models for study of human embryo implantation: choice of cell lines?
Biol Reprod. 2010 Feb;82(2):235-45. doi: 10.1095/biolreprod.109.077800. Epub 2009 Jul 1.
9
Cellular membrane potentials induced by alternating fields.
Biophys J. 1992 Dec;63(6):1632-42. doi: 10.1016/S0006-3495(92)81740-X.
10

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