Enhancing suspended cell transfection by inducing localized distribution of the membrane actin cortex before exposure to electromechanical stimulation.

IF 2 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Biotechnology Letters Pub Date : 2023-12-01 Epub Date: 2023-09-01 DOI:10.1007/s10529-023-03382-y
Wenjing Huang, Yibo Ma, Naotomo Tottori, Yoko Yamanishi
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引用次数: 0

Abstract

Objectives: During physical transfection, an electrical field or mechanical force is used to induce cell transfection. We tested if the disruption of a dense actin layer underneath the membrane of a suspended cell enhances cell transfection.

Results: A bubble generator was used to electromechanically stimulate suspended cells. To clarify the influence of the actin layer (the actin cortex) on cell transfection efficiency, we used an actin polymerization inhibitor (cytochalasin D) to disrupt the actin cortex before electromechanical stimulation. Without cytochalasin D treatment, signals from the overall actin cortex decreased after electromechanical stimulation. With cytochalasin D treatment, there was localized F-actin aggregation under static conditions. After electromechanical stimulation, there was a partial loss (localized disruption), but no overall disruption, of the actin cortex. With the pretreatment with cytochalasin D, the transfection efficiency of plasmids (4.7, 8.3, or 11 kbp) into NIH/3T3 or UMR-106 cells increased significantly after exposure to electromechanical stimulation.

Conclusions: Localized distribution of the actin cortex before exposure to electromechanical stimulation is crucial for inducing a partial loss of the cortex, which improves transfection efficiency and large plasmid delivery.

Abstract Image

在暴露于机电刺激前,通过诱导肌动蛋白皮层膜的局部分布来增强悬浮细胞转染。
目的:在物理转染过程中,使用电场或机械力来诱导细胞转染。我们测试了悬浮细胞膜下致密肌动蛋白层的破坏是否能增强细胞转染。结果:使用气泡发生器对悬浮细胞进行机电刺激。为了阐明肌动蛋白层(肌动蛋白皮层)对细胞转染效率的影响,我们使用肌动蛋白聚合抑制剂(细胞松弛素D)在机电刺激前破坏肌动蛋白皮层。在没有细胞松弛素D处理的情况下,来自整个肌动蛋白皮层的信号在机电刺激后减少。细胞松弛素D处理后,在静态条件下存在局部F-肌动蛋白聚集。在机电刺激后,肌动蛋白皮层出现部分损失(局部破坏),但没有整体破坏。在用细胞松弛素D预处理的情况下,暴露于机电刺激后,质粒(4.7、8.3或11kbp)转染NIH/3T3或UMR-106细胞的效率显著提高。结论:在暴露于机电刺激之前,肌动蛋白皮层的局部分布对于诱导皮层的部分缺失至关重要,这提高了转染效率和大质粒递送。
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来源期刊
Biotechnology Letters
Biotechnology Letters 工程技术-生物工程与应用微生物
CiteScore
5.90
自引率
3.70%
发文量
108
审稿时长
1.2 months
期刊介绍: Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.
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