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在气道上皮质细胞系中优化化学转染

Tony J F Guo¹, Wan Yi Liang², Gurpreet K Singhera²

¹Centre for Heart Lung Innovation, St. Paul's Hospital, Providence Healthcare Research Institute, University of British Columbia, 1081 Burrard St, Vancouver, BC, V6Z 1Y6, Canada. tony.guo@hli.ubc.ca.

BMC biotechnology

|January 24, 2025

概括

在呼吸道上皮模型中优化化学转染对于基因治疗研究至关重要. 甲胺3000显示高效率与最小的细胞影响,而其他试剂在细胞系显著变化.

科学领域:

细胞生物学细胞生物学
分子生物学分子生物学
基因治疗基因治疗

背景情况:

化学转染是研究气道表皮中的基因表达的一个关键方法.
它具有开发基因疗法的潜力.
这项研究评估了常见气道上皮细胞模型中的传染效率.

研究的目的:

用各种化学转化试剂来描述EX-EGFP-Lv105等离子体的转化效率.
为了比较不同传染试剂在三个气道上皮细胞系 (1HAEo-, 16HBE14o-和NCI-H292) 的疗效和细胞活力影响.
评估介质交换和素-EDTA预处理对转染结果的影响.

主要方法:

使用了六种常见的化学转染试剂:Lipofectamine 3000 (L3000),FuGENE HD,ViaFect,jetOPTIMUS,EndoFectin和酸.使用了六种常见的化学转染试剂:Lipofectamine 3000 (L3000),FuGENE HD,ViaFect,jetOPTIMUS,EndoFectin和酸.
将EX-EGFP-Lv105等离子体转化为1HAEo,16HBE14o和NCI-H292细胞系.
通过EGFP表达评估了转染效率,并在转染后48小时测量了细胞活力.
研究了6小时介质交换和氨酸-EDTA预处理对转染效率和生存能力的影响.

主要成果:

果胺3000 (L3000) 在1HAEo-细胞 (76.1%) 中表现出最高的转染效率,对细胞活力的影响最小.
与L3000相比,jetOPTIMUS在16HBE14o-细胞中显示出更高的效率 (64.6%),但可行性显著降低.
用素-EDTA进行预处理提高了1HAEo- (85.2%) 和16HBE14o- (62.6%) 细胞系的转化效率.

结论:

传染效率高度依赖于特定的气道上皮细胞系,使用的试剂和优化技术.
精心选择和优化转染条件对于改善非病毒基因输送在体外是必不可少的.
素-EDTA预处理提供了一个可行的策略,以提高某些气道上皮细胞模型的转染效率.

关键词:

气道上皮质气道上皮质化学转化过程化学转化过程优化优化优化优化

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