气道上皮细胞系化学转染的优化。

IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Tony J F Guo, Wan Yi Liang, Gurpreet K Singhera, Jasmine Memar Vaghri, Janice M Leung, Del R Dorscheid
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引用次数: 0

摘要

背景:化学转染是一种广泛应用于气道上皮研究的技术,可以研究基因表达的变化及其影响。此外,人们还在探索它在传递基因治疗方面的潜在应用。在这里,我们表征了表达egfp的质粒EX-EGFP-Lv105转染到三种常用的气道上皮模型细胞系(1HAEo-, 16HBE14o-和NCI-H292)的效率。结果:我们使用了六种常见和/或市售的化学成分不同的试剂:Lipofectamine 3000 (L3000)、FuGENE HD、ViaFect、jetOPTIMUS、EndoFectin和磷酸钙。使用L3000, 1HAEo-的转染效率高于16HBE14o-和NCI-H292 (1HAEo-: 76.1±3.2%,16HBE14o-: 35.5±1.2%,NCI-H292: 28.9±2.23%)。L3000的转染效率最高,对细胞活力的影响最小,归一化到对照组,转染后48小时,1HAEo-降低11.3±0.16%,16HBE14o-降低16.3±0.08%,NCI-H292降低17.5±0.09%。然而,jetOPTIMUS在1HAEo-中具有相似的转染效率(90.7±4.2%,p = 0.94),但显著降低了37.4±0.11%的细胞活力(p结论:转染效率可能因气道上皮细胞系、试剂和所使用的优化技术而异。考虑和优化细胞系和转染条件可能有助于改进体外非病毒遗传技术。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Optimization of chemical transfection in airway epithelial cell lines.

Background: Chemical transfection is a widely employed technique in airway epithelium research, enabling the study of gene expression changes and effects. Additionally, it has been explored for its potential application in delivering gene therapies. Here, we characterize the transfection efficiency of EX-EGFP-Lv105, an EGFP-expressing plasmid into three cell lines commonly used to model the airway epithelium (1HAEo-, 16HBE14o-, and NCI-H292).

Results: We used six common and/or commercially available reagents with varying chemical compositions: Lipofectamine 3000 (L3000), FuGENE HD, ViaFect, jetOPTIMUS, EndoFectin, and calcium phosphate. Using L3000, 1HAEo- exhibited the highest transfection efficiency compared to 16HBE14o- and NCI-H292 (1HAEo-: 76.1 ± 3.2%, 16HBE14o-: 35.5 ± 1.2%, NCI-H292: 28.9 ± 2.23%). L3000 yielded the greatest transfection efficiency with the lowest impact on cellular viability, normalized to control, with a 11.3 ± 0.16% reduction in 1HAEo-, 16.3 ± 0.08% reduction in 16HBE14o-, and 17.5 ± 0.09% reduction in NCI-H292 at 48-hour post-transfection. However, jetOPTIMUS had a similar transfection efficiency in 1HAEo- (90.7 ± 4.2%, p = 0.94), but had significantly reduced cellular viability of 37.4 ± 0.11% (p < 0.0001) compared to L3000. In 16HBE14o-, jetOPTIMUS yielded a significantly higher transfection efficiency compared to L3000 (64.6 ± 3.2%, p < 0.0001) but significantly reduced viability of 33.4 ± 0.09% (p < 0.0001) compared to L3000. In NCI-H292, jetOPTIMUS yielded a lower transfection efficiency (22.6 ± 1.2%) with a significant reduction in viability (28.3 ± 0.9%, p < 0.0001). Other reagents varied significantly in their efficiency and impact on cellular viability in other cell lines. Changing the transfection mixture-containing medium at 6-hour post-transfection did not improve transfection efficiency or viability. However, pre-treatment of cell cultures with two rinses of 0.25% trypsin-EDTA improved transfection efficiency in 1HAEo- (85.2 ± 1.1% vs. 71.3 ± 1.0%, p = 0.004) and 16HBE14o- (62.6 ± 4.3 vs. 35.5 ± 1.2, p = 0.003).

Conclusions: Transfection efficiencies can differ based on airway epithelial cell line, reagents, and optimization techniques used. Consideration and optimization of cell line and transfection conditions may be useful for improving nonviral genetic techniques in vitro.

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来源期刊
BMC Biotechnology
BMC Biotechnology 工程技术-生物工程与应用微生物
CiteScore
6.60
自引率
0.00%
发文量
34
审稿时长
2 months
期刊介绍: BMC Biotechnology is an open access, peer-reviewed journal that considers articles on the manipulation of biological macromolecules or organisms for use in experimental procedures, cellular and tissue engineering or in the pharmaceutical, agricultural biotechnology and allied industries.
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