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ScreenFect A:高效且低毒的脂质体,转染间充质干细胞

更新时间:2023-04-01   点击次数:461次

ScreenFect A: an efficient and low toxic liposome for gene delivery to mesenchymal stem cells

ScreenFect A:一种高效且低毒的脂质体,用于将基因传递到间充质干细胞

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摘要

间充质干细胞(MSCs)在包括组织工程和癌症治疗在内的各种治疗应用中具有巨大的前景。间充质干细胞的基因修饰可用于通过促进特定功能或将间充质干细胞转化为更有效的基因治疗工具来增强间充质干细胞的治疗效果。然而,转基因MSC的成功生成往往受到现有转染试剂转染效率差或毒性高的限制。在我们之前的研究中,我们使用硫醇-炔点击化学来开发新的脂质体载体,包括ScreenFect(A)  (® SF)(Li等人,2012)。本研究考察了SF对MSCs的转染性能。以Lipofectamine™ 2000(L2K)为对照,对转染效率、细胞活力和细胞DNA摄取进行了比较评估,结果表明SF在MSC转染方面优于L2K。血清的存在对SF或L2K的转染效率没有显著影响,但大大降低了L2K转染MSC的活力。与L2K相比,SF介导的转染效率更高,也与更好的细胞增殖相关。这些结果得到了DNA细胞内命运监测的支持,这证实了DNA从溶酶体的稳定运输和有效的核定位。SF的TGF-β1基因递送促进了成骨诱导条件下MSC成骨分化。作为SF脂质化对干细胞的研究,本研究强调了SF在向间充质干细胞和其他干细胞的基因传递中促进基于转基因干细胞的组织工程和其他治疗效果的有希望的作用。


Abstract

Mesenchymal stem cells (MSCs) hold great promise in variety of therapeutic applications including tissue engineering and cancer therapy. Genetic modification of MSCs can be used to enhance the therapeutic effect of MSCs by facilitating a specific function or by transforming MSCs into more effective gene therapy tools. However, the successful generation of genetically modified MSCs is often limited by the poor transfection efficiency or high toxicity of available transfection reagents. In our previous study, we used thiol-yne click chemistry to develop new liposomal vectors, including ScreenFect(®) A (SF) (Li et al., 2012). In this study, we investigated the transfection performance of SF on MSCs. A comparative evaluation of transfection efficiency, cell viability and cellular DNA uptake was performed using the Lipofectamine™ 2000 (L2K) as a control, and the results show that SF is superior to L2K for MSC transfection. The presence of serum did not significantly influence the transfection efficiency of either SF or L2K but greatly reduced the viability of MSC transfected by L2K. The higher efficiency of SF-mediated transfection compared to L2K was also correlated with better proliferation of cells. These results were supported by monitoring the intracellular fate of DNA, which confirmed stable transportation of DNA from lysosomes and efficient nuclear localization. TGF-β1 gene delivery by SF promoted MSC osteogenic differentiation in an osteogenic induction condition. As the first study of SF lipofection on stem cells, this study highlights a promising role of SF in gene delivery to MSCs as well as other stem cells to facilitate tissue engineering and other therapeutic effects based on genetically modified stem cells.


通过科研成果转化,ScreenFect已经成为一种商业转染试剂。德国ScreentFect联合靶点科技,将这种通过点击化学筛选出来的高效转染试剂带给中国广大的科研工作者。详情联系靶点科技。


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